Subcutaneous formulations of anti-CD38 antibodies and their uses

ABSTRACT

The present invention relates to subcutaneous formulations of anti-CD38 antibodies and their uses.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/460,754, filed Jul. 2, 2019, which is a continuation of U.S.application Ser. No. 15/340,290, filed Nov. 1, 2016, which claims thebenefit of U.S. Provisional Application No. 62/250,016, filed on Nov. 3,2015. The entire teachings of the above applications are incorporatedherein by reference.

INCORPORATION BY REFERENCE OF MATERIAL IN ASCII TEXT FILE

This application incorporates by reference the Sequence Listingcontained in the following ASCII text file being submitted concurrentlyherewith:

-   -   a) File name: 0148_2014_049_Sequence_Listing.txt; created Dec.        9, 2020, 26 KB in size.

FIELD OF THE INVENTION

The present invention relates to subcutaneous formulations of anti-CD38antibodies and their uses.

BACKGROUND OF THE INVENTION

CD38 is a multifunctional protein having function in receptor-mediatedadhesion and signaling as well as mediating calcium mobilization via itsecto-enzymatic activity, catalyzing formation of cyclic ADP-ribose(cADPR) and ADPR. CD38 mediates cytokine secretion and activation andproliferation of lymphocytes (Funaro et al., J Immunol 145:2390-6, 1990;Terhorst et al., Cell 771-80, 1981; Guse et al., Nature 398:70-3, 1999).CD38, via its NAD glycohydrolase activity, also regulates extracellularNAD⁺ levels, which have been implicated in modulating the regulatoryT-cell compartment (Adriouch et al., 14:1284-92, 2012; Chiarugi et al.,Nature Reviews 12:741-52, 2012). In addition to signaling via Ca²⁺, CD38signaling occurs via cross-talk with antigen-receptor complexes on T-and B-cells or other types of receptor complexes, e.g., MHC molecules,involving CD38 in several cellular responses, but also in switching andsecretion of IgG1. CD38 is expressed on various malignant cells.

Anti-CD38 antibodies are being developed for the treatment of multiplemyeloma and other heme malignancies. The antibodies are either injectedor infused via the intravenous (IV) route. The amount of antibody thatcan be administered via the intravenous route is limited by thephysico-chemical properties of the antibody, in particularly by itssolubility and stability in a suitable liquid formulation and by thevolume of the infusion fluid.

Therefore, there is a need for additional anti-CD38 antibodyformulations and pharmaceutical compositions.

SUMMARY OF THE INVENTION

The invention provides a pharmaceutical composition comprising ananti-CD38 antibody and a hyaluronidase.

The invention also provides a pharmaceutical composition comprising ananti-CD38 antibody and a hyaluronidase rHuPH20 having the amino acidsequence of SEQ ID NO: 22.

The invention also provides method of treating a cancer in a subject,comprising administering subcutaneously to the subject in need thereof apharmaceutical composition comprising an anti-CD38 antibody and ahyaluronidase for a time sufficient to treat the cancer.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering subcutaneously to asubject in need thereof the pharmaceutical composition of the inventionfor a time sufficient to treat the CD38-positive hematologicalmalignancy.

The invention also provides a method of treating a multiple myeloma,comprising administering subcutaneously to a subject in need thereof thepharmaceutical composition of the invention for a time sufficient totreat the multiple myeloma.

The invention also provides a unit dosage form, comprising

-   -   an anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of from about 1,200 mg to about        5,000 mg;    -   a hyaluronidase in an amount of from about 30,000 U to about        45,000 U;    -   histidine at a concentration of from about 5 mM to about 15 mM;    -   sorbitol at a concentration of from about 100 mM to about 300        mM;    -   PS-20 at a concentration of from about 0.01% w/v to about 0.04%        w/v; and    -   methionine at a concentration of from about 1 mg/mL to about 2        mg/mL, at a pH of about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the    -   VL of SEQ ID NO: 5 in an amount of about 1,800 mg;    -   a hyaluronidase in an amount of about 30,000 U;    -   histidine at a concentration of about 10 mM;    -   sorbitol at a concentration of about 300 mM;    -   PS-20 at a concentration of about 0.04% w/v; and    -   methionine at a concentration of about 1 mg/mL, at a pH of about        5.5.

The invention also provides a container comprising the unit dosage formof the invention.

The invention also provides a container comprising the pharmaceuticalcomposition of the invention.

DETAILED DESCRIPTION OF THE INVENTION

“CD38” refers to the human CD38 protein (synonyms: ADP-ribosyl cyclase1, cADPr hydrolase 1, cyclic ADP-ribose hydrolase 1). Human CD38 has anamino acid sequence shown in GenBank accession number NP 001766 and inSEQ ID NO: 1. It is well known that

CD38 is a single pass type II membrane protein with amino acid residues1-21 representing the cytosolic domain, amino acid residues 22-42representing the transmembrane domain, and residues 43-300 representingthe extracellular domain of CD38.

SEQ ID NO: 1 MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVVVLAVVVPRWRQQWSGPGTTKRFPETVLARCVKYTEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQRDMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEAACDVVHVMLNGSRSKIFDKNSTFGSVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCK NIYRPDKFLQCVKNPEDSSCTSEI

“Antibodies” is meant in a broad sense and includes immunoglobulinmolecules including monoclonal antibodies including murine, human,humanized and chimeric monoclonal antibodies, antigen-binding fragments,bispecific or multispecific antibodies, dimeric, tetrameric ormultimeric antibodies, single chain antibodies, domain antibodies andany other modified configuration of the immunoglobulin molecule thatcomprises an antigen binding site of the required specificity. “Fulllength antibodies” are comprised of two heavy (H) chains and two light(L) chains inter-connected by disulfide bonds as well as multimersthereof (for example IgM). Each heavy chain is comprised of a heavychain variable region (VH) and a heavy chain constant region (comprisedof domains CHL hinge CH2 and CH3). Each light chain is comprised of alight chain variable region (VL) and a light chain constant region (CL).The VH and the VL regions may be further subdivided into regions ofhypervariability, termed complementarity determining regions (CDR),interspersed with framework regions (FR). Each VH and VL is composed ofthree CDRs and four FR segments, arranged from amino-terminus tocarboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,CDR3, and FR4.

“Complementarity determining regions (CDR)” are “antigen binding sites”in an antibody. CDRs may be defined using various terms: (i)Complementarity Determining Regions (CDRs), three in the VH (HCDR1,HCDR2, HCDR3) and three in the VL (LCDR1, LCDR2, LCDR3) are based onsequence variability (Wu and Kabat, J Exp Med 132:211-50, 1970; Kabat etal., Sequences of Proteins of Immunological Interest, 5th Ed. PublicHealth Service, National Institutes of Health, Bethesda, Md., 1991).(ii) “Hypervariable regions”, “HVR”, or “HV”, three in the VH (H1, H2,H3) and three in the VL (L1, L2, L3) refer to the regions of an antibodyvariable domains which are hypervariable in structure as defined byChothia and Lesk (Chothia and Lesk, Mol Biol 196:901-17, 1987). TheInternational ImMunoGeneTics (IMGT) database (www_imgt_org) provides astandardized numbering and definition of antigen-binding sites. Thecorrespondence between CDRs, HVs and IMGT delineations is described inLefranc et al., Dev Comparat Immunol 27:55-77, 2003. The term “CDR”,“HCDR1”, “HCDR2”, “HCDR3”, “LCDR1”, “LCDR2” and “LCDR3” as used hereinincludes CDRs defined by any of the methods described supra, Kabat,Chothia or IMGT, unless otherwise explicitly stated in thespecification.

Immunoglobulins may be assigned to five major classes, IgA, IgD, IgE,IgG and IgM, depending on the heavy chain constant domain amino acidsequence. IgA and IgG are further sub-classified as the isotypes IgA₁,IgA₂, IgG₁, IgG₂, IgG₃ and IgG₄. Antibody light chains of any vertebratespecies can be assigned to one of two clearly distinct types, namelykappa (κ) and lambda (λ), based on the amino acid sequences of theirconstant domains.

“Antigen-binding fragment” refers to a portion of an immunoglobulinmolecule that retains the antigen binding properties of the parentalfull length antibody. Exemplary antigen-binding fragments are as heavychain complementarity determining regions (HCDR) 1, 2 and/or 3, lightchain complementarity determining regions (LCDR) 1, 2 and/or 3, a heavychain variable region (VH), or a light chain variable region (VL), Fab,F(ab′)2, Fd and Fv fragments as well as domain antibodies (dAb)consisting of either one VH domain or one VL domain. VH and VL domainsmay be linked together via a synthetic linker to form various types ofsingle chain antibody designs in which the VH/VL domains pairintramolecularly, or intermolecularly in those cases when the VH and VLdomains are expressed by separate chains, to form a monovalent antigenbinding site, such as single chain Fv (scFv) or diabody; described forexample in Int. Pat. Publ. No. WO1998/44001, Int. Pat. Publ. No.WO1988/01649; Int. Pat. Publ. No. WO1994/13804; Int. Pat. Publ. No.WO1992/01047.

“Monoclonal antibody” refers to an antibody population with single aminoacid composition in each heavy and each light chain, except for possiblewell known alterations such as removal of C-terminal lysine from theantibody heavy chain. Monoclonal antibodies typically bind one antigenicepitope, except that multispecific monoclonal antibodies bind two ormore distinct antigens or epitopes. Bispecific monoclonal antibodiesbind two distinct antigenic epitopes. Monoclonal antibodies may haveheterogeneous glycosylation within the antibody population. Monoclonalantibody may be monospecific or multispecific, or monovalent, bivalentor multivalent. A multispecific antibody, such as a bispecific antibodyor a trispecific antibody is included in the term monoclonal antibody.

“Isolated antibody” refers to an antibody or an antigen-binding fragmentthereof that is substantially free of other antibodies having differentantigenic specificities (e.g., an isolated antibody specifically bindinghuman CD38 is substantially free of antibodies that specifically bindantigens other than human CD38). In case of a bispecific antibody, thebispecific antibody specifically binds two antigens of interest, and issubstantially free of antibodies that specifically bind antigens otherthat the two antigens of interest. “Isolated antibody” encompassesantibodies that are isolated to a higher purity, such as antibodies thatare 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% pure.

“Humanized antibodies” refers to antibodies in which the antigen bindingsites are derived from non-human species and the variable regionframeworks are derived from human immunoglobulin sequences. Humanizedantibodies may include intentionally introduced mutations in theframework regions so that the framework may not be an exact copy ofexpressed human immunoglobulin or germline gene sequences.

“Human antibodies” refers to antibodies having heavy and light chainvariable regions in which both the framework and the antigen bindingsite are derived from sequences of human origin. If the antibodycontains a constant region or a portion of the constant region, theconstant region also is derived from sequences of human origin.

A human antibody comprises heavy or light chain variable regions thatare derived from sequences of human origin if the variable regions ofthe antibody are obtained from a system that uses human germlineimmunoglobulin or rearranged immunoglobulin genes. Such exemplarysystems are human immunoglobulin gene libraries displayed on phage, andtransgenic non-human animals such as mice or rats carrying humanimmunoglobulin loci as described herein. A human antibody typicallycontains amino acid differences when compared to the human germline orrearranged immunoglobulin sequences due to, for example naturallyoccurring somatic mutations, intentional introduction of substitutionsinto the framework or antigen binding site and amino acid changesintroduced during cloning and VDJ recombination in non-human animals.Typically, a human antibody is at least about 80%, 81%, 82%, 83%, 84%,85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99% or 100% identical in amino acid sequence to an amino acid sequenceencoded by a human germline or rearranged immunoglobulin gene. In somecases, a human antibody may contain consensus framework sequencesderived from human framework sequence analyses, for example as describedin Knappik et al., J Mol Biol 296:57-86, 2000, or synthetic HCDR3incorporated into human immunoglobulin gene libraries displayed onphage, for example as described in Shi et al., J Mol Biol 397:385-96,2010 and Int. Pat. Publ. No. WO2009/085462.

Antibodies in which antigen binding sites are derived from a non-humanspecies are not included in the definition of human antibody.

“Recombinant” includes antibodies and other proteins that are prepared,expressed, created or isolated by recombinant means.

“Epitope” refers to a portion of an antigen to which an antibodyspecifically binds. Epitopes typically consist of chemically active(such as polar, non-polar or hydrophobic) surface groupings of moietiessuch as amino acids or polysaccharide side chains and may have specificthree-dimensional structural characteristics, as well as specific chargecharacteristics. An epitope may be composed of contiguous and/ordiscontiguous amino acids that form a conformational spatial unit. For adiscontiguous epitope, amino acids from differing portions of the linearsequence of the antigen come in close proximity in 3-dimensional spacethrough the folding of the protein molecule.

“Multispecific” refers to an antibody that specifically binds at leasttwo distinct antigens or two distinct epitopes within the antigens, forexample three, four or five distinct antigens or epitopes.

“Bispecific” refers to an antibody that specifically binds two distinctantigens or two distinct epitopes within the same antigen. Thebispecific antibody may have cross-reactivity to other related antigensor can bind an epitope that is shared between two or more distinctantigens.

“Variant” refers to a polypeptide or a polynucleotide that differs froma reference polypeptide or a reference polynucleotide by one or moremodifications for example, substitutions, insertions or deletions.

“In combination with” means that two or more therapeutics areadministered to a subject together in a mixture, concurrently as singleagents or sequentially as single agents in any order.

“Pharmaceutical composition” refers to a product that results fromcombining an anti-CD38 antibody and a hyaluronidase and includes bothfixed and non-fixed combinations. Pharmaceutical composition typicallyincludes a pharmaceutically acceptable carrier. “Fixed combinations”refers to a single pharmaceutical composition comprising the anti-CD38antibody and the hyaluronidase administered simultaneously in the formof a single entity or dosage. “Non-fixed combination” refers to separatepharmaceutical compositions of the anti-CD38 antibody and thehyaluronidase or unit dosage forms administered as separate entitieseither simultaneously, concurrently or sequentially with no specificintervening time limits, wherein such administration provides effectivelevels of the two compounds in the body of the subject.

“Pharmaceutically acceptable carrier” refers to an ingredient in apharmaceutical composition, other than an active ingredient, which isnontoxic to a subject. A pharmaceutically acceptable carrier includes,but is not limited to, a buffer, excipient, stabilizer, or preservative.

“Treat” or “treatment” refers to therapeutic treatment wherein theobject is to slow down (lessen) an undesired physiological change ordisease, such as the development or spread of tumor or tumor cells, orto provide a beneficial or desired clinical outcome during treatment.Beneficial or desired clinical outcomes include alleviation of symptoms,diminishment of extent of disease, stabilized (i.e., not worsening)state of disease, delay or slowing of disease progression, lack ofmetastasis, amelioration or palliation of the disease state, andremission (whether partial or total), whether detectable orundetectable. “Treatment” may also mean prolonging survival as comparedto expected survival if a subject was not receiving treatment. Those inneed of treatment include those subjects already with the undesiredphysiological change or disease well as those subjects prone to have thephysiological change or disease.

“Therapeutically effective amount” refers to an amount effective, atdosages and for periods of time necessary, to achieve the desiredtherapeutic result. A therapeutically effective amount may varyaccording to factors such as the disease state, age, sex, and weight ofthe individual, and the ability of a therapeutic or a combination oftherapeutics to elicit a desired response in the individual. Exemplaryindicators of an effective therapeutic or combination of therapeuticsinclude, for example, improved well-being of the patient, reduction in atumor burden, arrested or slowed growth of a tumor, and/or absence ofmetastasis of cancer cells to other locations in the body.

“Inhibits growth” (e.g. referring to tumor cells) refers to a measurabledecrease in the tumor cell growth or tumor tissue in vitro or in vivowhen contacted with a therapeutic or a combination of therapeutics ordrugs, when compared to the growth of the same tumor cells or tumortissue in the absence of the therapeutic or the combination oftherapeutic drugs. Inhibition of growth of a tumor cell or tumor tissuein vitro or in vivo may be at least about 10%, 20%, 30%, 40%, 50%, 60%,70%, 80%, 90%, 99%, or 100%.

“CD38-positive hematological malignancy” refers to a hematologicalmalignancy characterized by the presence of tumor cells expressing CD38including leukemias, lymphomas and myeloma. Examples of suchCD38-positive hematological malignancies include precursor B-celllymphoblastic leukemia/lymphoma and B-cell non-Hodgkin's lymphoma, acutepromyelocytic leukemia, acute lymphoblastic leukemia and mature B-cellneoplasms, such as B-cell chronic lymphocytic leukemia (CLL)/smalllymphocytic lymphoma (SLL), B-cell acute lymphocytic leukemia, B-cellprolymphocytic leukemia, lymphoplasmacytic lymphoma, mantle celllymphoma (MCL), follicular lymphoma (FL), including low-grade,intermediate-grade and high-grade FL, cutaneous follicle centerlymphoma, marginal zone B-cell lymphoma (MALT type, nodal and splenictype), hairy cell leukemia, diffuse large B-cell lymphoma (DLBCL),Burkitt's lymphoma (BL), plasmacytoma, multiple myeloma, plasma cellleukemia, post-transplant lymphoproliferative disorder, light chainamyloidosis, Waldenstrom's macroglobulinemia, plasma cell leukemias andanaplastic large-cell lymphoma (ALCL).

1. “About” means within an acceptable error range for the particularvalue as determined by one of ordinary skill in the art, which willdepend in part on how the value is measured or determined, i.e., thelimitations of the measurement system. Unless explicitly statedotherwise within the Examples or elsewhere in the Specification in thecontext of a particular assay, result or embodiment, “about” meanswithin one standard deviation per the practice in the art, or a range ofup to 5%, whichever is larger.

Pharmaceutical Compositions

The invention provides a pharmaceutical composition comprising ananti-CD38 antibody and a hyaluronidase.

The pharmaceutical composition is useful for subcutaneous administrationof the anti-CD38 antibody to a subject in need of anti-CD38 antibodytherapy, such as a subject having a cancer, for example a CD38-positivehematological malignancy. Without wishing to be bound by any particulartheory, subcutaneous administration of the anti-CD38 antibody may havereduced infusion related reaction and achieve improved response rateswhen compared to the intravenous administration of the anti-CD38antibody.

In some embodiments, the pharmaceutical composition is a fixedcombination.

In some embodiments, the pharmaceutical composition is a non-fixedcombination.

In some embodiments, the pharmaceutical composition comprises from about1 mg/mL to about 180 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about10 mg/mL to about 180 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about20 mg/mL to about 160 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about20 mg/mL to about 140 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about20 mg/mL to about 120 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about40 mg/mL to about 120 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about60 mg/mL to about 120 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about80 mg/mL to about 120 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about100 mg/mL to about 120 mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about 1mg/mL, about 5 mg/mL, about 10 mg/mL, about 15 mg/mL, about 20 mg/mL,about 30 mg/mL, about 40 mg/mL, about 50 mg/mL, about 60 mg/mL, about 70mg/mL, about 80 mg/mL, about 90 mg/mL, about 100 mg/mL, about 110 mg/mL,about 120 mg/mL, about 130 mg/mL, about 140 mg/mL, about 150 mg/mL,about 160 mg/mL, about 170 mg/mL or about 180 mg/mL of the anti-CD38antibody.

In some embodiments, the pharmaceutical composition comprises about 20mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about 100mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about 120mg/mL of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about50 U/mL to about 5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about500 U/mL to about 5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about1,000 U/mL to about 5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about2,000 U/mL to about 5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about50 U/mL to about 2,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about500 U/mL to about 2,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about1,000 U/mL to about 2,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about 500U/mL, about 600 U/mL, about 700 U/mL, about 800 U/mL, about 900 U/mL,about 1,000 U/mL, about 1,100 U/mL, about 1,200 U/mL, about 1,300 U/mL,about 1,400 U/mL, about 1,500 U/mL, about 1,600 U/mL, about 1,700 U/mL,about 1,800 U/mL, about 1,900 U/mL, about 2,000 U/mL, about 2,100 U/mL,about 2,200 U/mL, about 2,300 U/mL, about 2,400 U/mL, about 2,500 U/mL,about 2,600 U/mL, about 2,700 U/mL, about 2,800 U/mL, about 2,900 U/mL,about 3,000 U/mL, about 3,100 U/mL, about 3,200 U/mL, about 3,300 U/mL,about 3,400 U/mL, about 3,500 U/mL, about 3,600 U/mL, about 3,700 U/mL,about 3,800 U/mL, about 3,900 U/mL, about 4,000 U/mL, about 4,100 U/mL,about 4,200 U/mL, about 4,300 U/mL, about 4,400 U/mL, about 4,500 U/mL,about 4,600 U/mL, about 4,700 U/mL, about 4,800 U/mL, about 4,900 U/mLor about 5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about 500U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about5,000 U/mL of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about1,200 mg to about 5,000 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about1,200 mg to about 2,400 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about1,200 mg to about 1,800 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about1,200 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about1,400 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about1,600 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about1,800 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about2,000 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about2,200 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about2,400 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about2,600 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about2,800 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about3,000 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about3,500 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about4,000 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about4,500 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises about5,000 mg of the anti-CD38 antibody.

In some embodiments, the pharmaceutical composition comprises from about750 U to about 75,000 U of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about7,500 U to about 45,000 U of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises from about30,000 U to about 45,000 U of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about7,500 U, about 8,000 U, about 8,500 U, about 9,000 U, about 10,000 U,about 15,000 U, about 20,000 U, about 21,000 U, about 22,000 U, about23,000 U, about 24,000 U, about 25,000 U, about 26,000 U, about 27,000U, about 28,000 U, about 29,000 U, about 30,000 U, about 31,000 U, about32,000 U, about 33,000 U, about 34,000 U, about 35,000 U, about 36,000U, about 37,000 U, about 38,000 U, about 39,000 U, about 40,000 U, about41,000 U, about 42,000 U, about 43,000 U, about 44,000 U, about 45,000U, about 46,000 U, about 47,000 U, about 48,000 U, about 49,000 U, about50,000 U, about 55,000 U, about 60,000 U, about 65,000 U, about 70,000 Uor about 75,000 U of the hyaluronidase.

In some embodiments, the pharmaceutical composition comprises about5,000 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about5,000 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about3,000 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about3,000 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,800 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,800 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,600 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,600 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,400 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,400 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,200 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,200 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,000 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about2,000 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about1,800 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about1,800 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about1,600 mg of the anti-CD38 antibody and about 30,000 U of thehyaluronidase.

In some embodiments, the pharmaceutical composition comprises about1,600 mg of the anti-CD38 antibody and about 45,000 U of thehyaluronidase.

In some embodiments, the hyaluronidase is rHuPH20 having the amino acidsequence of SEQ ID NO: 22.

rHuPH20 is a recombinant hyaluronidase (HYLENEX® recombinant) and isdescribed in Int. Pat. Publ. No. WO2004/078140.

Hyaluronidase is an enzyme that degrades hyaluronic acid (EC 3.2.1.35)and lowers the viscosity of hyaluronan in the extracellular matrix,thereby increasing tissue permeability.

Enzymatic activity of hyaluronidase, including rHuPH20 can be defined byunits per mL (U/mL) or by total enzyme activity in a particularformulation (U).

The standard definition for one unit (U) of enzyme activity is theamount of enzyme that catalyzes the reaction of 1 nmol of substrate perminute.

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition competes for binding to CD38 with an antibody comprising aheavy chain variable region (VH) of SEQ ID NO: 4 and a light chainvariable region (VL) of SEQ ID NO: 5.

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition binds at least to the region SKRNIQFSCKNIYR (SEQ ID NO: 2)and the region EKVQTLEAWVIHGG (SEQ ID NO: 3) of human CD38 (SEQ ID NO:1).

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition comprises a heavy chain complementarity determining region 1(HCDR1), a HCDR2, a HCDR3, a light chain complementarity determiningregion 1 (LCDR1), a LCDR2 and a LCDR3 of SEQ ID NOs: 6, 7 and 8, 9, 10and 11, respectively.

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition comprises a heavy chain variable region (VH) that is 95%,96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQID NO: 4 and a light chain variable region (VL) that is 95%, 96%, 97%,98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 5.

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition comprises the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5.

In some embodiments, the anti-CD38 antibody in the pharmaceuticalcomposition comprises a heavy chain of SEQ ID NO: 12 and a light chainof SEQ ID NO: 13.

SEQ ID NO: 2 SKRNIQFSCKNIYR SEQ ID NO: 3 EKVQTLEAWVIEGG SEQ ID NO: 4EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSS SEQ ID NO: 5EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPTF GQGTKVEIK SEQ ID NO: 6SFAMS SEQ ID NO: 7 AISGSGGGTYYADSVKG SEQ ID NO: 8 DKILWFGEPVFDYSEQ ID NO: 9 RASQSVSSYLA SEQ ID NO: 10 DASNRAT SEQ ID NO: 11 QQRSNWPPTFSEQ ID NO: 12 EVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFAMSWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKSEQ ID NO: 13 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV THQGLSSPVTKSFNRGEC

Other exemplary anti-CD38 antibodies that may be used in thepharmaceutical compositions and the methods of the invention are:

mAb003 comprising the VH and the VL sequences of SEQ ID NOs: 14 and 15,respectively and described in U.S. Pat. No. 7,829,693. The VH and the VLof mAb003 may be expressed as IgG1/K;

mAb024 comprising the VH and the VL sequences of SEQ ID NOs: 16 and 17,respectively, described in U.S. Pat. No. 7,829,693. The VH and the VL ofmAb024 may be expressed as IgG1/K;

MOR-202 (MOR-03087) comprising the VH and the VL sequences of SEQ IDNOs: 18 and 19, respectively, described in U.S. Pat. No. 8,088,896. TheVH and the VL of MOR-202 may be expressed as IgG1/K; or

Isatuximab; comprising the VH and the VL sequences of SEQ ID NOs: 20 and21, respectively, described in U.S. Pat. No. 8,153,765. The VH and theVL of Isatuximab may be expressed as Ig

SEQ ID NO: 14 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAFSWVRQAPGQGLEWMGRVIPFLGIANSAQKFQGRVTITADKSTSTAYMDLSSLRSEDTAVYYCARDDIAALGPFDYWGQGTLVTVSSAS SEQ ID NO: 15DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPRTF GQGTKVEIKSEQ ID NO: 16 EVQLVQSGAEVKKPGESLKISCKGSGYSFSNYWIGWVRQMPGKGLEWMGIIYPHDSDARYSPSFQGQVTFSADKSISTAYLQWSSLKASDTAMYYCARHVGWGSRYWYFDLWGRGTLVTVSS SEQ ID NO: 17EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPGLLIYDASNRASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGG GTKVEIK SEQ ID NO: 18QVQLVESGGGLVQPGGSLRLSCAASGFTFSSYYMNWVRQAPGKGLEWVSGISGDPSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR DLPLVYTGFAYWGQGTLVTVSSSEQ ID NO: 19 DIELTQPPSVSVAPGQTARISCSGDNLRHYYVYWYQQKPGQAPVLVIYGDSKRPSGIPERFSGSNSGNTATLTISGTQAEDEADYYCQTYTGGASLVF GGGTKLTVLGQSEQ ID NO 20: QVQLVQSGAEVAKPGTSVKLSCKASGYTFTDYWMQWVKQRPGQGLEWIGTIYPGDGDTGYAQKFQGKATLTADKSSKTVYMHLSSLASEDSAVYYCARGD YYGSNSLDYWGQGTSVTVSSSEQ ID NO: 21: DIVMTQSHLSMSTSLGDPVSITCKASQDVSTVVAWYQQKPGQSPRRLIYSASYRYIGVPDRFTGSGAGTDFTFTISSVQAEDLAVYYCQQHYSPPYTFGG GTKLEIK

Other exemplary anti-CD38 antibodies that may be used in thepharmaceutical compositions of the invention are those described in Int.Pat. Publ. No. WO05/103083, Intl. Pat. Publ. No. WO06/125640, Intl. Pat.Publ. No. WO07/042309, Intl. Pat. Publ. No. WO08/047242 or Intl. Pat.Publ. No. WO14/178820.

An exemplary anti-CD38 antibody that may be used in the pharmaceuticalcompositions of the invention is daratumumab. Daratumumab comprises theheavy chain variable region (VH) and the light chain variable region(VL) amino acid sequences shown in SEQ ID NOs: 4 and 5, respectively,the HCDR1, the HCDR2 and the HCDR3 of SEQ ID NOs: 6, 7 and 8,respectively, the LCDR1, the LCDR2 and the LCDR3 of SEQ ID NOs: 9, 10and 11, respectively, and is of IgG1/K subtype and described in U.S.Pat. No. 7,829,693. Daratumumab heavy chain amino acid sequence is shownin SEQ ID NO: 12 and light chain amino acid sequence shown in SEQ ID NO:13.

The invention also provides a pharmaceutical composition comprising ananti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the VL of SEQID NO: 5 and a hyaluronidase rHuPH20 of SEQ ID NO: 22, wherein theanti-CD38 antibody concentration in the pharmaceutical composition isabout 20 mg/mL.

The invention also provides a pharmaceutical composition comprising ananti-CD38 antibody comprising the HCDR1, the HCDR2 and the HCDR3 of SEQID NOs: 6, 7 and 8, respectively, and the LCDR1, the LCDR2 and the LCDR3of SEQ ID NOs: 9, 10 and 11, respectively and the hyaluronidase rHuPH20of SEQ ID NO: 22, wherein the anti-CD38 antibody concentration in thepharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingbetween about 1,200 mg-1,800 mg of the anti-CD38 antibody comprising theVH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5, and between about 30,000U-45,000 U of the hyaluronidase rHuPH20 of SEQ ID NO: 22, wherein theanti-CD38 antibody concentration in the pharmaceutical composition isabout 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,800 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 30,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,800 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 45,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,600 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 30,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,600 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 45,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,200 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 30,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

The invention also provides a pharmaceutical composition comprisingabout 1,200 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5, and about 45,000 U of the hyaluronidaserHuPH20 of SEQ ID NO: 22, wherein the anti-CD38 antibody concentrationin the pharmaceutical composition is about 20 mg/mL.

SEQ ID NO: 22 MGVLKFKHIFFRSFVKSSGVSQIVFTFLLIPCCLTLNFRAPPVIPNVPFLWAWNAPSEFCLGKFDEPLDMSLFSFIGSPRINATGQGVTIFYVDRLGYYPYIDSITGVTVNGGIPQKISLQDHLDKAKKDITFYMPVDNLGMAVIDWEEWRPTWARNWKPKDVYKNRSIELVQQQNVQLSLTEATEKAKQEFEKAGKDFLVETIKLGKLLRPNHLWGYYLFPDCYNHHYKKPGYNGSCFNVEIKRNDDLSWLWNESTALYPSIYLNTQQSPVAATLYVRNRVREAIRVSKIPDAKSPLPVFAYTRIVFTDQVLKFLSQDELVYTFGETVALGASGIVIWGTLSIMRSMKSCLLLDNYMETILNPYIINVTLAAKMCSQVLCQEQGVCIRKNWNSSDYLHLNPDNFAIQLEKGGKFTVRGKPTLEDLEQFSEKFYCSCYSTLSCKEKADVKDTDAVDVCIADGVCIDAFLKPPMETEEPQIFYNASPSTLSATMFIVSILFLIISSVASL

Anti-CD38 antibodies used in the pharmaceutical compositions of theinvention, may also be selected de novo from, e.g., a phage displaylibrary, where the phage is engineered to express human immunoglobulinsor portions thereof such as Fabs, single chain antibodies (scFv), orunpaired or paired antibody variable regions (Knappik et al., J Mol Biol296:57-86, 2000; Krebs et al., J Immunol Meth 254:67-84, 2001; Vaughanet al., Nature Biotechnology 14:309-314, 1996; Sheets et al., PITAS(USA) 95:6157-6162, 1998; Hoogenboom and Winter, J Mol Biol 227:381,1991; Marks et al., J Mol Biol 222:581, 1991). CD38 binding variabledomains may be isolated from e.g., phage display libraries expressingantibody heavy and light chain variable regions as fusion proteins withbacteriophage pIX coat protein as described in Shi et al., J. Mol. Biol.397:385-96, 2010 and Intl. Pat. Publ. No. WO09/085462). The antibodylibraries may be screened for binding to human CD38 extracellulardomain, the obtained positive clones further characterized, Fabsisolated from the clone lysates, and subsequently cloned as full lengthantibodies. Such phage display methods for isolating human antibodiesare established in the art. See for example: U.S. Pat. Nos. 5,223,409,5,403,484, 5,571,698, 5,427,908, 5,580,717, 5,969,108, 6,172,197,5,885,793, 6,521,404, 6,544,731, 6,555,313, 6,582,915, and 6,593,081.

Antibodies may be evaluated for their competition with a referenceantibody such as the anti-CD38 antibody comprising the VH of SEQ ID NO:4 and the VL of SEQ ID NO: 5 for binding to CD38 using known in vitromethods. In an exemplary method, CHO cells recombinantly expressing CD38may be incubated with unlabeled reference antibody for 15 min at 4° C.,followed by incubation with an excess of fluorescently labeled testantibody for 45 min at 4° C. After washing in PBS/BSA, fluorescence maybe measured by flow cytometry using standard methods. In anotherexemplary method, extracellular portion of human CD38 may be coated onthe surface of an ELISA plate. Excess of unlabeled reference antibodymay be added for about 15 minutes and subsequently biotinylated testantibodies may be added. After washes in PBS/Tween, binding of the testbiotinylated antibody may be detected using horseradish peroxidase(HRP)-conjugated streptavidine and the signal detected using standardmethods. It is readily apparent that in the competition assays, thereference antibody may be labelled and the test antibody unlabeled. Thetest antibody competes with the reference antibody when the referenceantibody inhibits binding of the test antibody, or the test antibodyinhibits binding of the reference antibody by at least 80%, for example81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 100%. The epitope of the test antibody mayfurther be defined for example by peptide mapping or hydrogen/deuteriumprotection assays using known methods, or by crystal structuredetermination.

Antibodies binding to the region SKRNIQFSCKNIYR (SEQ ID NO: 2) and theregion EKVQTLEAWVIHGG (SEQ ID NO: 3) of human CD38 (SEQ ID NO: 1) may begenerated for example by immunizing mice with peptides having the aminoacid sequences shown in SEQ ID NOs: 2 and 3 using standard methods andthose described herein, and characterizing the obtained antibodies forbinding to the peptides using for example ELISA or mutagenesis studies.

The invention also provides a pharmaceutical composition comprising ananti-CD38 antibody comprising the HCDR1, the HCDR2, the HCDR3, theLCDR1, the LCDR2, and the LCDR3 sequences of:

-   -   a. the VH of SEQ ID NO: 14 and the VL of SEQ ID NO: 15;    -   b. the VH of SEQ ID NO: 16 and the VL of SEQ ID NO: 17;    -   c. the VH of SEQ ID NO: 18 and the VL of SEQ ID NO: 19; or    -   d. the VH of SEQ ID NO: 20 and the VL of SEQ ID NO: 21, and the        hyaluronidase rHuPH20 of SEQ ID NO: 22.

The invention also provides a pharmaceutical composition comprising ananti-CD38 antibody comprising

-   -   a. the VH of SEQ ID NO: 14 and the VL of SEQ ID NO: 15;    -   b. the VH of SEQ ID NO: 16 and the VL of SEQ ID NO: 17;    -   c. the VH of SEQ ID NO: 18 and the VL of SEQ ID NO: 19; or    -   d. the VH of SEQ ID NO: 20 and the VL of SEQ ID NO: 21, and the        hyaluronidase rHuPH20 of SEQ ID NO: 22.

The pharmaceutical compositions of the invention further comprise apharmaceutically acceptable carrier. Exemplary pharmaceuticallyacceptable carriers are solvents, dispersion media, coatings,antibacterial and antifungal agents, isotonic and absorption delayingagents, and the like that are physiologically compatible, such as salts,buffers, antioxidants, saccharides, aqueous or non-aqueous carriers,preservatives, wetting agents, surfactants or emulsifying agents, orcombinations thereof.

Exemplary buffers that may be used are acetic acid, citric acid, formicacid, succinic acid, phosphoric acid, carbonic acid, malic acid,aspartic acid, histidine, boric acid, Tris buffers, HEPPSO and HEPES.

Exemplary antioxidants that may be used are ascorbic acid, methionine,cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodiumsulfite, lecithin, citric acid, ethylenediamine tetraacetic acid (EDTA),sorbitol and tartaric acid.

Exemplary amino acids that may be used are histidine, isoleucine,methionine, glycine, arginine, lysine, L-leucine, tri-leucine, alanine,glutamic acid, L-threonine, and 2-phenylamine.

Exemplary surfactants that may be used are polysorbates (e.g.,polysorbate-20 or polysorbate-80); polyoxamers (e.g., poloxamer 188);Triton; sodium octyl glycoside; lauryl-, myristyl-, linoleyl-, orstearyl-sulfobetaine; lauryl-, myristyl-, linoleyl- orstearyl-sarcosine; linoleyl-, myristyl-, or cetyl-betaine;lauroamidopropyl-, cocamidopropyl-, linoleamidopropyl-,myristamidopropyl-, palmidopropyl-, or isostearamidopropyl-betaine(e.g., lauroamidopropyl); myristamidopropyl-, palmidopropyl-, orisostearamidopropyl-dimethylamine; sodium methyl cocoyl-, or disodiummethyl oleyl-taurate; and the MONAQUA™ series (Mona Industries, Inc.,Paterson, N.J.), polyethyl glycol, polypropyl glycol, and copolymers ofethylene and propylene glycol (e.g., PLURONICS™, PF68, etc).

Exemplary preservatives that may be used are phenol, m-cresol, p-cresol,o-cresol, chlorocresol, benzyl alcohol, phenylmercuric nitrite,phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride,alkylparaben (methyl, ethyl, propyl, butyl and the like), benzalkoniumchloride, benzethonium chloride, sodium dehydroacetate and thimerosal,or mixtures thereof.

Exemplary saccharides that may be used are monosaccharides,disaccharides, trisaccharides, polysaccharides, sugar alcohols, reducingsugars, nonreducing sugars such as glucose, sucrose, trehalose, lactose,fructose, maltose, dextran, glycerin, dextran, erythritol, glycerol,arabitol, sylitol, sorbitol, mannitol, mellibiose, melezitose,raffinose, mannotriose, stachyose, maltose, lactulose, maltulose,glucitol, maltitol, lactitol or iso-maltulose.

Exemplary salts that may be used are acid addition salts and baseaddition salts. Acid addition salts include those derived from nontoxicinorganic acids, such as hydrochloric, nitric, phosphoric, sulfuric,hydrobromic, hydroiodic, phosphorous and the like, as well as fromnontoxic organic acids such as aliphatic mono- and dicarboxylic acids,phenyl-substituted alkanoic acids, hydroxy alkanoic acids, aromaticacids, aliphatic and aromatic sulfonic acids and the like. Base additionsalts include those derived from alkaline earth metals, such as sodium,potassium, magnesium, calcium and the like, as well as from nontoxicorganic amines, such as N,N′-dibenzylethylenediamine, N-methylglucamine,chloroprocaine, choline, diethanolamine, ethylenediamine, procaine andthe like. An exemplary salt is sodium chloride.

The amounts of pharmaceutically acceptable carrier(s) in thepharmaceutical compositions may be determined experimentally based onthe activities of the carrier(s) and the desired characteristics of theformulation, such as stability and/or minimal oxidation.

In some embodiments, the pharmaceutical composition comprises aceticacid.

In some embodiments, the pharmaceutical composition comprises aceticacid at a concentration of from about 1 mM to about 50 mM.

In some embodiments, the pharmaceutical composition comprises aceticacid at a concentration of from about 10 mM to about 40 mM.

In some embodiments, the pharmaceutical composition comprises aceticacid at a concentration of about 10 mM, about 15 mM, about 20 mM, about25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM or about 50mM.

In some embodiments, the pharmaceutical composition comprises aceticacid at a concentration of about 25 mM.

In some embodiments, the pharmaceutical composition comprises sodiumchloride (NaCl).

In some embodiments, the pharmaceutical composition comprises NaCl at aconcentration of from about 20 mM to about 100 mM.

In some embodiments, the pharmaceutical composition comprises NaCl at aconcentration of from about 40 mM to about 80 mM.

In some embodiments, the pharmaceutical composition comprises NaCl at aconcentration of about 20 mM, about 25 mM, about 30 mM, about 35 mM,about 40 mM, about 45 mM, about 50 mM, about 55 mM, about 60 mM, about65 mM, about 70 mM, about 75 mM, about 80 mM, about 85 mM, about 90 mM,about 95 mM or about 100 mM.

In some embodiments, the pharmaceutical composition comprises NaCl at aconcentration of about 60 mM.

In some embodiments, the pharmaceutical composition comprisessaccharide.

In some embodiments, saccharide is sucrose.

In some embodiments, saccharide is sorbitol.

In some embodiments, saccharide is mannitol.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 50 mM to about 500 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 50 mM to about 450 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 50 mM to about 400 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 50 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 100 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of from about 100 mM to about 300 mM.

In some embodiments, the pharmaceutical composition comprises saccharideat a concentration of about 100 mM, about 110 mM, about 120 mM, about130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about230 mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about280 mM, about 290 mM, about 300 mM, about 310 mM, about 320 mM, about330 mM, about 340 mM, about 350 mM, about 360 mM, about 370 mM, about380 mM, about 390 mM, about 400 mM, about 410 mM, about 420 mM, about430 mM, about 440 mM, about 450 mM, about 460 mM, about 470 mM, about480 mM, about 490 mM or about 500 mM.

In some embodiments, the pharmaceutical composition comprises mannitol.

In some embodiments, the pharmaceutical composition comprises mannitolat a concentration of from about 100 mM to about 180 mM.

In some embodiments, the pharmaceutical composition comprises mannitolat a concentration of from about 120 mM to about 160 mM.

In some embodiments, the pharmaceutical composition comprises mannitolat a concentration of about 100 mM, about 105 mM, about 110 mM, about115 mM, about 120 mM, about 125 mM, about 130 mM, about 135 mM, about140 mM, about 145 mM, about 150 mM, about 155 mM, about 160 mM, about165 mM, about 170 mM, about 175 mM or about 180 mM.

In some embodiments, the pharmaceutical composition comprises mannitolat a concentration of about 140 mM.

In some embodiments, the pharmaceutical composition comprisespolysorbate.

In some embodiments, the pharmaceutical composition comprisespolysorbate-20 (PS-20).

In some embodiments, the pharmaceutical composition comprisespolysorbate-20 (PS-20) at a concentration of from about 0.01% w/v toabout 0.1% w/v.

In some embodiments, the pharmaceutical composition comprisespolysorbate-20 (PS-20) at a concentration of from about 0.01% w/v toabout 0.08% w/v.

In some embodiments, the pharmaceutical composition comprisespolysorbate-20 (PS-20) at a concentration of from about 0.01% w/v toabout 0.04% w/v.

In some embodiments, the pharmaceutical composition comprisespolysorbate-20 (PS-20) at a concentration of about 0.01% w/v, 0.02% w/v,0.03% w/v, 0.04% w/v, 0.05% w/v, 0.06% w/v, 0.07% w/v, 0.08% w/v, 0.09%w/v or 0.1% w/v.

The invention also provides a pharmaceutical composition comprising

-   -   from about 20 mg/mL to about 120 mg/mL of the anti-CD38 antibody        comprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in        about 25 mM acetic acid, about 60 mM sodium chloride, about 140        mM mannitol and about 0.04% w/v polysorbate-20 (PS-20); at pH        about 5.5; and    -   about from 30,000 U to about 45,000 U of the hyaluronidase in 10        mM L-Histidine, 130 mM NaCl, 10 mM L-Methionine, 0.02%        Polysorbate 80, pH 6.5.

In some embodiments, the hyaluronidase is rHuPH20 (SEQ ID NO: 22).

In some embodiments, the pharmaceutical composition is a non-fixedcombination.

The invention also provides a pharmaceutical composition comprising

-   -   about 20 mg/mL of the anti-CD38 antibody comprising the VH of        SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in about 25 mM acetic        acid, about 60 mM sodium chloride, about 140 mM mannitol and        about 0.04% w/v polysorbate-20 (PS-20); at pH about 5.5; and        about 30,000 U of the hyaluronidase in 10 mM L-Histidine, 130 mM        NaCl, 10 mM L-Methionine, 0.02% Polysorbate 80, pH 6.5.

In some embodiments, the hyaluronidase is rHuPH20 (SEQ ID NO: 22).

In some embodiments, the pharmaceutical composition is a non-fixedcombination.

The invention also provides a pharmaceutical composition comprising

-   -   about 20 mg/mL of the anti-CD38 antibody comprising the VH of        SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in about 25 mM acetic        acid, about 60 mM sodium chloride, about 140 mM mannitol and        about 0.04% w/v polysorbate-20 (PS-20); at pH about 5.5; and    -   about 45,000 U of the hyaluronidase in 10 mM L-Histidine, 130 mM        NaCl, 10 mM L-Methionine, 0.02% Polysorbate 80, pH 6.5.

In some embodiments, the hyaluronidase is rHuPH20 (SEQ ID NO: 22).

In some embodiments, the pharmaceutical composition is a non-fixedcombination.

In some embodiments, the pharmaceutical composition comprises histidine.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 1 mM to about 50 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 5 mM to about 50 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 5 mM to about 30 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 5 mM to about 20 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 5 mM to about 15 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of from about 5 mM to about 10 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of about 1 mM, about 2 mM, about 3 mM, about 4 mM,about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM,about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM,about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about27 mM, about 28 mM, about 29 mM, about 30 mM, about 31 mM, about 32 mM,about 33 mM, about 34 mM, about 35 mM, about 36 mM, about 37 mM, about38 mM, about 39 mM, about 40 mM, about 41 mM, about 42 mM, about 43 mM,about 44 mM, about 45 mM, about 46 mM, about 47 mM, about 48 mM, about49 mM or about 50 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of about 5 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of about 10 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of about 15 mM.

In some embodiments, the pharmaceutical composition comprises histidineat a concentration of about 20 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 50 mM to about 500 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 50 mM to about 450 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 50 mM to about 400 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 50 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 100 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of from about 100 mM to about 300 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 100 mM, about 110 mM, about 120 mM, about130 mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about230 mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about280 mM, about 290 mM, about 300 mM, about 310 mM, about 320 mM, about330 mM, about 340 mM, about 350 mM, about 360 mM, about 370 mM, about380 mM, about 390 mM, about 400 mM, about 410 mM, about 420 mM, about430 mM, about 440 mM, about 450 mM, about 460 mM, about 470 mM, about480 mM, about 490 mM or about 500 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 50 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 100 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 150 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 200 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 250 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 300 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 350 mM.

In some embodiments, the pharmaceutical composition comprises sorbitolat a concentration of about 400 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 50 mM to about 500 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 50 mM to about 450 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 50 mM to about 400 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 50 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 100 mM to about 350 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of from about 100 mM to about 200 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 100 mM, about 110 mM, about 120 mM, about 130mM, about 140 mM, about 150 mM, about 160 mM, about 170 mM, about 180mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230mM, about 240 mM, about 250 mM, about 260 mM, about 270 mM, about 280mM, about 290 mM, about 300 mM, about 310 mM, about 320 mM, about 330mM, about 340 mM, about 350 mM, about 360 mM, about 370 mM, about 380mM, about 390 mM, about 400 mM, about 410 mM, about 420 mM, about 430mM, about 440 mM, about 450 mM, about 460 mM, about 470 mM, about 480mM, about 490 mM or about 500 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 50 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 100 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 150 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 200 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 250 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 300 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 350 mM.

In some embodiments, the pharmaceutical composition comprises sucrose ata concentration of about 400 mM.

In some embodiments, the pharmaceutical composition comprisesmethionine.

In some embodiments, the pharmaceutical composition comprises methionineat a concentration of from about 0.1 mg/mL to about 5 mg/mL.

In some embodiments, the pharmaceutical composition comprises methionineat a concentration of from about 0.1 mg/mL to about 2.5 mg/mL.

In some embodiments, the pharmaceutical composition comprises methionineat a concentration of from about 1 mg/mL to about 2 mg/mL.

In some embodiments, the pharmaceutical composition comprises methionineat a concentration of about 0.5 mg/mL, about 1 mg/mL, about 1.1 mg/mL,about 1.2 mg/mL, about 1.3 mg/mL, about 1.4 mg/mL, about 1.5 mg/mL,about 1.6 mg/mL, about 1/7 mg/mL, about 1.8 mg/mL, about 1.9 mg/mL,about 2.0 mg/mL, about 2.1 mg/mL, about 2.2 mg/mL, about ⅔ mg/mL, about2.4 mg/mL, about 2.5 mg/mL, about 2.6 mg/mL, about 2.7 mg/mL, about 2.8mg/mL, about 2.9 mg/mL, about 3 mg/mL, about 3.5 mg/mL, about 4 mg/mL,about 4.5 mg/mL or about 5 mg/mL.

In some embodiments, the pharmaceutical composition is at pH 5.0 to 6.0.

In some embodiments, the pharmaceutical composition is at pH 5.3 to 5.8.

In some embodiments, the pharmaceutical composition is at pH 5.5.

In some embodiments, the pharmaceutical composition is at pH 5.6.

The invention also provides a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase    -   from about 5 mM to about 50 mM histidine; and    -   from about 50 mM to about 400 mM sorbitol.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase from        about 5 mM to about 50 mM histidine;    -   from about 50 mM to about 400 mM sorbitol;    -   from about 0.01% w/v to about 0.1% PS-20; and    -   from about 0.1 mg/mL to about 2.5 mg/mL methionine.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a pharmaceutical composition comprising

-   -   from about 100 mg/mL to about 120 mg/mL of the anti-CD38        antibody;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase;    -   about 10 mM histidine; and    -   from about 100 mM to about 300 mM sorbitol.

In some embodiments, the hyaluronidase is rHuPH20.

In some embodiments, the pharmaceutical composition further comprisesfrom

-   about 0.01% w/v to about 0.04% w/v PS-20.

In some embodiments, the pharmaceutical composition further comprisesfrom about 1 mg/mL to about 2 mg/mL methionine.

In some embodiments, the pharmaceutical composition further comprisesfrom about 100 mM to about 200 mM sucrose.

In some embodiments, the anti-CD38 antibody comprises

-   -   the HCDR1, the HCDR2, the HCDR3, the LCDR1, the LCDR2 and the        LCDR3 of SEQ ID NOs: 6, 7, 8, 9, 10 and 11, respectively;    -   the VH and the VL of SEQ ID NOs: 4 and 5, respectively; and/or    -   the heavy chain and the light chain of SEQ ID NOs: 12 and 13,        respectively.

In some embodiments, the anti-CD38 antibody comprises

-   -   the VH and the VL of SEQ ID NOs: 14 and 15, respectively;    -   the VH and the VL of SEQ ID NOs: 16 and 17, respectively;    -   the VH and the VL of SEQ ID NOs: 18 and 19, respectively; or    -   the VH and the VL of SEQ ID NOs: 20 and 21, respectively;

In some embodiments, the hyaluronidase comprises rHuPH20 (SEQ ID NO: 22)

The invention also provides a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody        comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase    -   from about 5 mM to about 50 mM histidine; and    -   from about 50 mM to about 400 mM sorbitol.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody        comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively;    -   from about 50 U/mL to about 5,000 U/mL hyaluronidase    -   from about 5 mM to about 50 mM histidine;    -   from about 50 mM to about 400 mM sorbitol;    -   from about 0.01% w/v to about 0.1% PS-20; and    -   from about 0.1 mg/mL to about 2.5 mg/mL methionine.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a pharmaceutical composition comprising

-   -   from about 100 mg/mL to about 120 mg/mL of the anti-CD38        antibody comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively;    -   from about 50 U/mL to about 5,000 U/mL rHuPH20;    -   about 10 mM histidine;    -   from about 100 mM to about 300 mM sorbitol;    -   from about 0.01% w/v to about 0.04% w/v PS-20; and    -   from about 1 mg/mL to about 2 mg/mL methionine.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 2 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 120 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 2,000 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.6.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol; and    -   about 2 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.01% w/v PS-20; and    -   about 2 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.02% w/v PS-20; and    -   about 2 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.06% w/v PS-20; and    -   about 2 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 50 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 2,000 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.5.

The invention also provides a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 5,000 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.5.

In some embodiments, the pharmaceutical composition is a fixedcombination.

The formulations to be used for in vivo administration are generallysterile. Sterility may be readily accomplished, e.g., by filtrationthrough sterile filtration membranes.

The pharmaceutical compositions of the invention may be prepared byknown methods. For example, the pharmaceutical compositions may beprepared, e.g., by dissolving, suspending or emulsifying the anti-CD38antibody in a sterile aqueous medium or an oily medium conventionallyused for injections.

Administration

The pharmaceutical compositions of the invention may be administered asa non-fixed combination.

The pharmaceutical compositions of the invention may also beadministered as a fixed combination, e.g., as a unit dosage form (ordosage unit form). Fixed combinations may be advantageous for ease ofadministration and uniformity of dosage.

The invention also provides a unit dosage form, comprising the anti-CD38antibody comprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 inan amount of from about 1,200 mg to about 5,000 mg and rHuPH20 in anamount of from about 30,000 U to about 75,000 U.

The invention also provides a unit dosage form, comprising the anti-CD38antibody comprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 inan amount of from about 1,200 mg to about 4,000 mg and rHuPH20 in anamount of from about 30,000 U to about 75,000 U.

The invention also provides a unit dosage form, comprising the anti-CD38antibody comprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 inan amount of from about 1,200 mg to about 2,400 mg and rHuPH20 in anamount of from about 30,000 U to about 45,000 U.

The invention also provides a unit dosage form, comprising the anti-CD38antibody comprising the VH of

-   SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in an amount of from about    1,200 mg to about 1,800 mg and rHuPH20 in an amount of from about    30,000 U to about 45,000 U.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of from about 1,200 mg to about        5,000 mg;    -   rHuPH20 in an amount of from about 30,000 U to about 75,000 U;        histidine at a concentration of from about 5 mM to about 15 mM;        sorbitol at a concentration of from about 100 mM to about 300        mM; PS-20 at a concentration of from about 0.01% w/v to about        0.04% w/v; and    -   methionine at a concentration of from about 1 mg/mL to about 2        mg/mL, at a pH of about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of about 1,200 mg to about 2,400        mg;    -   rHuPH20 in an amount of from about 30,000 U to about 45,000 U;    -   histidine at a concentration of about 10 mM;    -   sorbitol at a concentration of about 300 mM;    -   PS-20 at a concentration of about 0.04% w/v; and    -   methionine at a concentration of from about 1 mg/mL; at a pH of        about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of about 1,200 mg to about 1,800        mg;    -   rHuPH20 in an amount of from about 30,000 U to about 45,000 U;    -   histidine at a concentration of about 10 mM;    -   sorbitol at a concentration of about 300 mM;    -   PS-20 at a concentration of about 0.04% w/v; and    -   methionine at a concentration of from about 1 mg/mL; at a pH of        about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of from about 1,200 mg to about        1,800 mg;    -   rHuPH20 in an amount of from about 30,000 U to about 45,000 U;    -   histidine at a concentration of from about 5 mM to about 15 mM;    -   sorbitol at a concentration of from about 100 mM to about 300        mM;    -   PS-20 at a concentration of from about 0.01% w/v to about 0.04%        w/v; and    -   methionine at a concentration of from about 1 mg/mL to about 2        mg/mL, at a pH of about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of about 1,800 mg;    -   rHuPH20 in an amount of from about 30,000 U;    -   histidine at a concentration of about 10 mM;    -   sorbitol at a concentration of about 300 mM;    -   PS-20 at a concentration of about 0.04% w/v; and    -   methionine at a concentration of from about 1 mg/mL; at a pH of        about 5.5.

The invention also provides a unit dosage form, comprising

-   -   the anti-CD38 antibody comprising the VH of SEQ ID NO: 4 and the        VL of SEQ ID NO: 5 in an amount of about 1,800 mg;    -   rHuPH20 in an amount of from about 45,000 U;    -   histidine at a concentration of about 10 mM;    -   sorbitol at a concentration of about 300 mM;    -   PS-20 at a concentration of about 0.04% w/v; and    -   methionine at a concentration of from about 1 mg/mL; at a pH of        about 5.5.

The pharmaceutical composition of the invention may be administered in atotal volume of about 80 mL, 90 mL, 100 mL, 110 mL or 120 mL.

The pharmaceutical composition of the invention may be administered in atotal volume of about 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL,17 mL, 18 mL, 19 mL, 20 mL, 25 mL, 30 mL, 35 mL, 40 mL, 45 mL, 50 mL, 55mL, 60 mL, 65 mL, 70 mL, 75 mL, 80 mL, 85 mL, 90 mL, 95 mL, 100 mL, 105mL, 110 mL, 115 mL or 120 mL.

The pharmaceutical composition of the invention may be administered in atotal volume of about 10 mL.

The pharmaceutical composition of the invention may be administered in atotal volume of about 15 mL.

The pharmaceutical composition of the invention may be administered in atotal volume of about 20 mL.

The total volume of administration may be typically smaller for thefixed combinations when compared to the non-fixed combinations.

The invention also provides a container comprising the pharmaceuticalcomposition of the invention.

The invention also provides a container comprising the unit dosage formof the invention.

The container may be a vial, a cartridge, a syringe, a prefilled syringeor a disposable pen.

The administration of the pharmaceutical compositions of the inventionmay be repeated after one day, two days, three days, four days, fivedays, six days, one week, two weeks, three weeks, four weeks, fiveweeks, six weeks, seven weeks, two months, three months, four months,five months, six months or longer. Repeated courses of treatment arealso possible, as is chronic administration. The repeated administrationmay be at the same dose or at a different dose. For example, thepharmaceutical compositions of the invention may be administered onceweekly for eight weeks, followed by once in two weeks for 16 weeks,followed by once in four weeks.

The pharmaceutical composition of the invention may be administeredsubcutaneously.

The pharmaceutical composition of the invention may be administeredsubcutaneously to the abdominal region.

Subcutaneous administration may be accomplished using a device. Thedevice may be a syringe, a prefilled syringe, an auto-injector, eitherdisposable or reusable, a pen injector, a patch injector, a wearableinjector or an ambulatory syringe infusion pump with subcutaneousinfusion sets.

For non-fixed combinations, 20 mg/mL anti-CD38 antibody in 25 mM sodiumacetate, 60 mM sodium chloride, 140 mM D-mannitol, 0.04% polysorbate 20,pH 5.5 may be mixed with 1 mg/mL (75-150 kU/mL) rHuPH20 in 10 mML-Histidine, 130 mM NaCl, 10 mM L-Methionine, 0.02% polysorbate-80, pH6.5 prior to administration of the mixture to a subject.

The pharmaceutical compositions of the invention may also beadministered prophylactically in order to reduce the risk of developingcancer, delay the onset of the occurrence of an event in cancerprogression, and/or reduce the risk of recurrence when a cancer is inremission. This may be especially useful in patients wherein it isdifficult to locate a tumor that is known to be present due to otherbiological factors.

Methods of Treatment

The invention also provides a method of treating a cancer, comprisingadministering to a subject in need thereof the pharmaceuticalcomposition of the invention for a time sufficient to treat the cancer.

In some embodiments, the cancer is a CD38-positive hematologicalmalignancy.

In some embodiments, the CD38-positive hematological malignancy ismultiple myeloma.

In some embodiments, the CD38-positive hematological malignancy isdiffuse large B-cell lymphoma (DLBCL).

In some embodiments, the CD38-positive hematological malignancy isnon-Hodgkin's lymphoma.

In some embodiments, the CD38-positive hematological malignancy is acutelymphoblastic leukemia (ALL).

In some embodiments, the CD38-positive hematological malignancy isfollicular lymphoma (FL).

In some embodiments, the CD38-positive hematological malignancy isBurkitt's lymphoma (BL).

In some embodiments, the CD38-positive hematological malignancy ismantle cell lymphoma (MCL).

In some embodiments, the CD38-positive hematological malignancy is lightchain amyloidosis (AL).

In some embodiments, the CD38-positive hematological malignancy ismultiple myeloma, acute lymphoblastic leukemia (ALL), non-Hodgkin'slymphoma, diffuse large B-cell lymphoma (DLBCL), Burkitt's lymphoma(BL), follicular lymphoma (FL) or mantle-cell lymphoma (MCL).

Examples of B-cell non-Hodgkin's lymphomas are lymphomatoidgranulomatosis, primary effusion lymphoma, intravascular large B-celllymphoma, mediastinal large B-cell lymphoma, heavy chain diseases(including γ, μ, and a disease), lymphomas induced by therapy withimmunosuppressive agents, such as cyclosporine-induced lymphoma, andmethotrexate-induced lymphoma.

In some embodiments, the cancer is a solid tumor.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising an anti-CD38 antibodyand a hyaluronidase subcutaneously for a time sufficient to treat theCD38-positive hematological malignancy, wherein the anti-CD38 antibodyconcentration in the pharmaceutical composition is about 20 mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising the anti-CD38 antibodycomprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 and thehyaluronidase rHuPH20 of SEQ ID NO: 22 subcutaneously for a timesufficient to treat the CD38-positive hematological malignancy, whereinthe anti-CD38 antibody concentration in the pharmaceutical compositionis about 20 mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising between about 1,200mg-1,800 mg of the anti-CD38 antibody comprising the VH of SEQ ID NO: 4and the VL of SEQ ID NO: 5, and between about 30,000 U-45,000 U of thehyaluronidase rHuPH20 of SEQ ID NO: 22 for a time sufficient to treatthe CD38-positive hematological malignancy, wherein the anti-CD38antibody concentration in the pharmaceutical composition is about 20mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising between about 1,200 mgand about 1,800 mg of the anti-CD38 antibody comprising the VH of SEQ IDNO: 4 and the VL of SEQ ID NO: 5, and about 30,000 U of thehyaluronidase rHuPH20 of SEQ ID NO: 22 for a time sufficient to treatthe CD38-positive hematological malignancy, wherein the anti-CD38antibody concentration in the pharmaceutical composition is about 20mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising between about 1,200 mgand about 1,800 mg of the anti-CD38 antibody comprising the VH of SEQ IDNO: 4 and the VL of SEQ ID NO: 5, and about 45,000 U of thehyaluronidase rHuPH20 of SEQ ID NO: 22 for a time sufficient to treatthe CD38-positive hematological malignancy, wherein the anti-CD38antibody concentration in the pharmaceutical composition is about 20mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof between about 1,600 mg of the anti-CD38 antibody comprising theVH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5, and about 30,000 U of thehyaluronidase rHuPH20 of SEQ ID NO: 22 for a time sufficient to treatthe CD38-positive hematological malignancy, wherein the anti-CD38antibody concentration in the pharmaceutical composition is about 20mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof about 1,600 mg of the anti-CD38 antibody comprising the VH ofSEQ ID NO: 4 and the VL of SEQ ID NO: 5, and about 45,000 U of thehyaluronidase rHuPH20 of SEQ ID NO: 22 for a time sufficient to treatthe CD38-positive hematological malignancy, wherein the anti-CD38antibody concentration in the pharmaceutical composition is about 20mg/mL.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising the anti-CD38 antibodycomprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 and thehyaluronidase, wherein the pharmaceutical composition is a non-fixedcombination.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   from about 20 mg/mL to about 120 mg/mL of the anti-CD38 antibody        comprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in        about 25 mM acetic acid, about 60 mM sodium chloride, about 140        mannitol and about 0.04% w/v polysorbate-20 (PS-20); at pH about        5.5; and    -   about from 30,000 U to about 45,000 U of the hyaluronidase in 10        mM L-histidine, 130 mM NaCl, 10 mM L-methionine, 0.02%        Polysorbate-80, pH 6.5.

In some embodiments, the hyaluronidase is rHuPH20.

In some embodiments, the pharmaceutical composition is a non-fixedcombination.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   about 20 mg/mL of the anti-CD38 antibody comprising the VH of        SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in about 25 mM acetic        acid, about 60 mM sodium chloride, about 140 mannitol and about        0.04% w/v polysorbate-20 (PS-20); at pH about 5.5; and    -   about 30,000 U of the hyaluronidase in 10 mM L-histidine, 130 mM        NaCl, 10 mM L-methionine, 0.02% Polysorbate-80, pH 6.5.    -   In some embodiments, the hyaluronidase is rHuPH20.    -   In some embodiments, the pharmaceutical composition is a        non-fixed combination.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   about 20 mg/mL of the anti-CD38 antibody comprising the VH of        SEQ ID NO: 4 and the VL of SEQ ID NO: 5 in about 25 mM acetic        acid, about 60 mM sodium chloride, about 140 mannitol and about        0.04% w/v polysorbate-20 (PS-20); at pH about 5.5; and    -   about 45,000 U of the hyaluronidase in 10 mM L-histidine, 130 mM        NaCl, 10 mM L-methionine, 0.02% Polysorbate-80, pH 6.5.    -   In some embodiments, the hyaluronidase is rHuPH20.    -   In some embodiments, the pharmaceutical composition is a        non-fixed combination.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising the anti-CD38 antibodycomprising the VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 and thehyaluronidase, wherein the pharmaceutical composition is a fixedcombination.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody        comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase    -   from about 5 mM to about 50 mM histidine; and    -   from about 50 mM to about 400 mM sorbitol.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   from about 1 mg/mL to about 180 mg/mL of the anti-CD38 antibody        comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively;    -   from about 50 U/mL to about 5,000 U/mL of the hyaluronidase    -   from about 5 mM to about 50 mM histidine;    -   from about 50 mM to about 400 mM sorbitol;    -   from about 0.01% w/v to about 0.1% PS-20; and    -   from about 0.1 mg/mL to about 2.5 mg/mL methionine.    -   In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   from about 100 mg/mL to about 120 mg/mL of the anti-CD38        antibody comprising the VH and the VL of SEQ ID NOs: 4 and 5,        respectively; from about 50 U/mL to about 5,000 U/mL of the        hyaluronidase;    -   about 10 mM histidine;    -   from about 100 mM to about 300 mM sorbitol;    -   from about 0.01% w/v to about 0.04% w/v PS-20; and    -   from about 1 mg/mL to about 2 mg/mL methionine.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   about 100 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 500 U/mL of the hyaluronidase;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 2 mg/mL methionine at pH about 5.5.

In some embodiments, the hyaluronidase is rHuPH20.

The invention also provides a method of treating a CD38-positivehematological malignancy, comprising administering to a subject in needthereof a pharmaceutical composition comprising

-   -   about 120 mg/mL of the anti-CD38 antibody comprising the VH and        the VL of SEQ ID NOs: 4 and 5, respectively;    -   about 2,000 U/mL rHuPH20;    -   about 10 mM histidine;    -   about 300 mM sorbitol;    -   about 0.04% w/v PS-20; and    -   about 1 mg/mL methionine; at pH about 5.6.

In some embodiments, the hyaluronidase is rHuPH20.

The anti-CD38 antibodies in the pharmaceutical compositions of theinvention may induce killing of CD38-expressing tumor cells byantibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependentcellular phagocytosis (ADCP), complement dependent cytotoxicity (CDC),apoptosis, or modulation of CD38 enzymatic activity. The anti-CD38antibodies in the pharmaceutical compositions of the invention may alsomediate anti-tumor efficacy by their immunomodulatory effects byinducing CD4+ and CD8⁺ T cell proliferation, and/or by relievinginhibition of inflammatory responses mediated by myeloid-derivedsuppressor cells (MDSCs) and regulatory T cells (Tregs).

“Antibody-dependent cellular cytotoxicity”, “antibody-dependentcell-mediated cytotoxicity” or “ADCC” is a mechanism for inducing celldeath that depends upon the interaction of antibody-coated target cellswith effector cells possessing lytic activity, such as natural killercells, monocytes, macrophages and neutrophils via Fc gamma receptors(FcγR) expressed on effector cells. For example, NK cells expressFcγRIIIa, whereas monocytes express FcγRI, FcγRII and FcvRIIIa. Death ofthe antibody-coated target cell, such as CD38-expressing cells, occursas a result of effector cell activity through the secretion of membranepore-forming proteins and proteases. To assess ADCC activity of anantibody that specifically binds CD38, the antibody may be added toCD38-expressing cells in combination with immune effector cells, whichmay be activated by the antigen antibody complexes resulting incytolysis of the target cell. Cytolysis is generally detected by therelease of label (e.g. radioactive substrates, fluorescent dyes ornatural intracellular proteins) from the lysed cells. Exemplary effectorcells for such assays include peripheral blood mononuclear cells (PBMC)and NK cells. Exemplary target cells include Tregs or MDSCs expressingCD38. In an exemplary assay, target cells are labeled with 20 μCi of⁵¹Cr for 2 hours and washed extensively. Cell concentration of thetarget cells may be adjusted to 1×10⁶ cells/ml, and anti-CD38 antibodiesat various concentrations are added. Assays are started by adding targetcells at an effector:target cell ratio of 40:1. After incubation for 3hr at 37° C. assays are stopped by centrifugation, and ⁵¹Cr release fromlysed cells are measured in a scintillation counter. Percentage ofcellular cytotoxicity may be calculated as % maximal lysis which may beinduced by adding 3% perchloric acid to target cells.

“Antibody-dependent cellular phagocytosis” (“ADCP”) refers to amechanism of elimination of antibody-coated target cells byinternalization by phagocytic cells, such as macrophages or dendriticcells. ADCP may be evaluated by using Tregs or MDSCs expressing CD38 astarget cells engineered to express GFP or other labeled molecule.Effctor:target cell ratio may be for example 4:1. Effector cells may beincubated with target cells for 4 hours with or without anti-CD38antibody. After incubation, cells may be detached using accutase.Macrophages may be identified with anti-CD11b and anti-CD14 antibodiescoupled to a fluorescent label, and percent phagocytosis may bedetermined based on % GFP fluorescent in the CD11⁺CD14⁺ macrophagesusing standard methods.

“Complement-dependent cytotoxicity”, or “CDC”, refers to a mechanism forinducing cell death in which an Fc effector domain of a target-boundantibody binds and activates complement component Clq which in turnactivates the complement cascade leading to target cell death.Activation of complement may also result in deposition of complementcomponents on the target cell surface that facilitate ADCC by bindingcomplement receptors (e.g., CR3) on leukocytes.

The ability of monoclonal antibodies to induce ADCC may be enhanced byengineering their oligosaccharide component. Human IgG1 or IgG3 areN-glycosylated at Asn297 with the majority of the glycans in thewell-known biantennary G0, G0F, G1, G1F, G2 or G2F forms. Antibodiesproduced by non-engineered CHO cells typically have a glycan fucosecontent of about at least 85%. The removal of the core fucose from thebiantennary complex-type oligosaccharides attached to the Fc regionsenhances the ADCC of antibodies via improved FcγRIIIa binding withoutaltering antigen binding or CDC activity. Such mAbs may be achievedusing different methods reported to lead to the successful expression ofrelatively high defucosylated antibodies bearing the biantennarycomplex-type of Fc oligosaccharides such as control of cultureosmolality (Konno et al., Cytotechnology 64:249-65, 2012), applicationof a variant CHO line Lec13 as the host cell line (Shields et al., JBiol Chem 277:26733-26740, 2002), application of a variant CHO line EB66as the host cell line (Olivier et al., MAbs; 2(4), 2010; Epub ahead ofprint; PMID:20562582), application of a rat hybridoma cell line YB2/0 asthe host cell line (Shinkawa et al., J Biol Chem 278:3466-3473, 2003),introduction of small interfering RNA specifically against the a1,6-fucosyltrasferase (FUT8) gene (Mori et al., Biotechnol Bioeng88:901-908, 2004), or coexpression ofβ-1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II ora potent alpha-mannosidase I inhibitor, kifunensine (Ferrara et al., JBiol Chem 281:5032-5036, 2006, Ferrara et al., Biotechnol Bioeng93:851-861, 2006; Xhou et al., Biotechnol Bioeng 99:652-65, 2008). ADCCelicited by anti-CD38 antibodies used in the methods of the invention,and in some embodiments of each and every one of the numberedembodiments listed below, may also be enhanced by certain substitutionsin the antibody Fc. Exemplary substitutions are for examplesubstitutions at amino acid positions 256, 290, 298, 312, 356, 330, 333,334, 360, 378 or 430 (residue numbering according to the EU index) asdescribed in U.S. Pat. No. 6,737,056.

In some embodiments, the anti-CD38 antibody comprises a substitution inthe antibody Fc.

In some embodiments, the anti-CD38 antibody comprises a substitution inthe antibody Fc at amino acid positions 256, 290, 298, 312, 356, 330,333, 334, 360, 378 or 430 (residue numbering according to the EU index).

In some embodiments, the anti-CD38 antibody has a biantennary glycanstructure with fucose content of about between 0% to about 15%, forexample 15%, 14%, 13%, 12%, 11% 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%or 0%.

In some embodiments, the anti-CD38 antibody has a biantennary glycanstructure with fucose content of about 50%, 40%, 45%, 40%, 35%, 30%,25%, 20%, 15%, 14%, 13%, 12%, 11% 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,1% or 0%

Substitutions in the Fc and reduced fucose content may enhance the ADCCactivity of the antibody that specifically binds CD38.

“Fucose content” means the amount of the fucose monosaccharide withinthe sugar chain at Asn297. The relative amount of fucose is thepercentage of fucose-containing structures related to allglycostructures. These may be characterized and quantified by multiplemethods, for example: 1) using MALDI-TOF of N-glycosidase F treatedsample (e.g. complex, hybrid and oligo- and high-mannose structures) asdescribed in Intl. Pat. Publ. No. WO2008/077546; 2) by enzymatic releaseof the Asn297 glycans with subsequent derivatization anddetection/quantitation by HPLC (UPLC) with fluorescence detection and/orHPLC-MS (UPLC-MS); 3) intact protein analysis of the native or reducedmAb, with or without treatment of the Asn297 glycans with Endo S orother enzyme that cleaves between the first and the second GlcNAcmonosaccharides, leaving the fucose attached to the first GlcNAc; 4)digestion of the mAb to constituent peptides by enzymatic digestion(e.g., trypsin or endopeptidase Lys-C), and subsequent separation,detection and quantitation by HPLC-MS (UPLC-MS) or 5) separation of themAb oligosaccharides from the mAb protein by specific enzymaticdeglycosylation with PNGase F at Asn 297. The oligosaccharides releasedmay be labeled with a fluorophore, separated and identified by variouscomplementary techniques which allow: fine characterization of theglycan structures by matrix-assisted laser desorption ionization (MALDI)mass spectrometry by comparison of the experimental masses with thetheoretical masses, determination of the degree of sialylation by ionexchange HPLC (GlycoSep C), separation and quantification of theoligosacharride forms according to hydrophilicity criteria bynormal-phase HPLC (GlycoSep N), and separation and quantification of theoligosaccharides by high performance capillary electrophoresis-laserinduced fluorescence (HPCE-LIF).

“Low fucose” or “low fucose content” as used herein refers to antibodieswith fucose content of about 0%-15%.

“Normal fucose” or “normal fucose content” as used herein refers toantibodies with fucose content of about over 50%, typically about over60%, 70%, 80% or over 85%.

In the methods described herein, and in some embodiments of each andevery one of the numbered embodiments listed below, the anti-CD38antibody is of IgG1, IgG2, IgG3 or IgG4 isotype.

Antibodies that are substantially identical to the antibody comprisingthe VH of SEQ ID NO: 4 and the VL of SEQ ID NO: 5 may be used in themethods of the invention. The term “substantially identical” as usedherein means that the two antibody VH or VL amino acid sequences beingcompared are identical or have “insubstantial differences”.Insubstantial differences are substitutions of 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, or 15 amino acids in an antibody heavy chain orlight chain that do not adversely affect antibody properties. Percentidentity may be determined for example by pairwise alignment using thedefault settings of the AlignX module of Vector NTI v.9.0.0 (Invitrogen,Carlsbad, Calif.). The protein sequences of the present invention may beused as a query sequence to perform a search against public or patentdatabases to, for example, identify related sequences. Exemplaryprograms used to perform such searches are the XBLAST or BLASTP programs(www_ncbi_nlm/nih_gov), or the GenomeQuest™ (GenomeQuest, Westborough,Mass.) suite using the default settings. Exemplary substitutions thatmay be made to the anti-CD38 antibodies used in the methods of theinvention are for example conservative substitutions with an amino acidhaving similar charge, hydrophobic, or stereochemical characteristics.Conservative substitutions may also be made to improve antibodyproperties, for example stability or affinity, or to improve antibodyeffector functions. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15amino acid substitutions may be made for example to the heavy or thelight chain of the anti-CD38 antibody. Furthermore, any native residuein the heavy or light chain may also be substituted with alanine, as hasbeen previously described for alanine scanning mutagenesis (MacLennan etal., Acta Physiol Scand Suppl 643:55-67, 1998; Sasaki et al., AdvBiophys 35:1-24, 1998). Desired amino acid substitutions may bedetermined by those skilled in the art at the time such substitutionsare desired. Amino acid substitutions may be done for example by PCRmutagenesis (U.S. Pat. No. 4,683,195). Libraries of variants may begenerated using well known methods, for example using random (NNK) ornon-random codons, for example DVK codons, which encode 11 amino acids(Ala, Cys, Asp, Glu, Gly, Lys, Asn, Arg, Ser, Tyr, Trp) and screeningthe libraries for variants with desired properties. The generatedvariants may be tested for their binding to CD38, their ability toinduce ADCC, ADCP or apoptosis, or modulate CD38 enzymatic activity invitro using methods described herein.

In some embodiments, the anti-CD38 antibody may bind human CD38 with arange of affinities (K_(D)). In one embodiment according to theinvention, and in some embodiments of each and every one of the numberedembodiments listed below, the anti-CD38 antibody binds to CD38 with highaffinity, for example, with a K_(D) equal to or less than about 10⁻⁷ M,such as but not limited to, 1-9.9 (or any range or value therein, suchas 1, 2, 3, 4, 5, 6, 7, 8, or 9)×10⁻⁸M, 10⁻⁹M, 10⁻¹⁰ M, 10⁻¹¹M, 10⁻¹²M,10⁻¹³M, 10⁻¹⁴M, 10⁻¹⁵ M or any range or value therein, as determined bysurface plasmon resonance or the Kinexa method, as practiced by those ofskill in the art. One exemplary affinity is equal to or less than 1×10⁻⁸M. Another exemplary affinity is equal to or less than 1×10⁻⁹ M.

In some embodiments, the anti-CD38 antibody is a bispecific antibody.The VL and/or the VH regions of the existing anti-CD38 antibodies or theVL and VH regions identified de novo as described herein may beengineered into bispecific full length antibodies. Such bispecificantibodies may be made by modulating the CH3 interactions between themonospecific antibody heavy chains to form bispecific antibodies usingtechnologies such as those described in U.S. Pat. No. 7,695,936; Intl.Pat. Publ. No. WO04/111233; U.S. Pat. Publ. No. US2010/0015133; U.S.Pat. Publ. No. US2007/0287170; Intl. Pat. Publ. No. WO2008/119353; U.S.Pat. Publ. No. US2009/0182127; U.S. Pat. Publ. No. US2010/0286374; U.S.Pat. Publ. No. US2011/0123532; Intl. Pat. Publ. No. WO2011/131746; Int.Pat. Publ. No. WO2011/143545; or U.S. Pat. Publ. No. US2012/0149876.Additional bispecific structures into which the VL and/or the VH regionsof the antibodies of the invention may be incorporated are for exampleDual Variable Domain Immunoglobulins (Inlt. Pat. Publ. No.WO2009/134776), or structures that include various dimerization domainsto connect the two antibody arms with different specificity, such asleucine zipper or collagen dimerization domains (Int. Pat. Publ. No.WO2012/022811, U.S. Pat. Nos. 5,932,448; 6,833,441).

For example, bispecific antibodies may be generated in vitro in acell-free environment by introducing asymmetrical mutations in the CH3regions of two monospecific homodimeric antibodies and forming thebispecific heterodimeric antibody from two parental monospecifichomodimeric antibodies in reducing conditions to allow disulfide bondisomerization according to methods described in Intl. Pat. Publ. No.WO2011/131746. In the methods, the first monospecific bivalent antibody(e.g., anti-CD38 antibody) and the second monospecific bivalent antibodyare engineered to have certain substitutions at the CH3 domain thatpromote heterodimer stability; the antibodies are incubated togetherunder reducing conditions sufficient to allow the cysteines in the hingeregion to undergo disulfide bond isomerization; thereby generating thebispecific antibody by Fab arm exchange. The incubation conditions mayoptimally be restored to non-reducing. Exemplary reducing agents thatmay be used are 2-mercaptoethylamine (2-MEA), dithiothreitol (DTT),dithioerythritol (DTE), glutathione, tris(2-carboxyethyl)phosphine(TCEP), L-cysteine and beta-mercaptoethanol, preferably a reducing agentselected from the group consisting of: 2-mercaptoethylamine,dithiothreitol and tris(2-carboxyethyl)phosphine. For example,incubation for at least 90 min at a temperature of at least 20° C. inthe presence of at least 25 mM 2-MEA or in the presence of at least 0.5mM dithiothreitol at a pH of from 5-8, for example at pH of 7.0 or at pHof 7.4 may be used.

Exemplary CH3 mutations that may be used in a first heavy chain and in asecond heavy chain of the bispecific antibody are K409R and/or F405L.

The methods of the invention may be used to treat an animal patientbelonging to any classification. Examples of such animals includemammals such as humans, rodents, dogs, cats and farm animals.

Combination Therapies

The pharmaceutical compositions of the invention may be administered incombination with a second therapeutic agent, or combinations thereof.

The second therapeutic agent may be melphalan, mechlorethamine, thioepa,chlorambucil, carmustine (BSNU), lomustine (CCNU), cyclophosphamide,busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC),procarbazine, mitomycin C, cisplatin and other platinum derivatives,such as carboplatin, thalidomide or a thalidomide analog, lenalidomideor CC4047, a proteasome inhibitor, such as bortezomib or vinca alkaloid,such as vincristine or an anthracycline, such as doxorubicin.

In some embodiments, the second therapeutic agent is a proteasomeinhibitor.

In some embodiments, the proteasome inhibitor is bortezomib, carfilzomibor ixazomib.

In some embodiments, the second therapeutic agent is an alkylatingagent.

In some embodiments, the alkylating agent is busulfan, cyclophosphamide,bendamustine, chlorambucli, carboplatin, cisplatin, temozolomide,melphalan, busulfan, bendamustine, carmustine, lomustine, dacarbazine,oxaliplatin, ifosfamide, mechlorethamine, thiotepa, trabectedin orstreptozocin.

In some embodiments, the second therapeutic agent is a glutamic acidderivative.

In some embodiments, the glutamic acid derivative is Revlimid®(lenalidomide), thalidomide or Pomalyst® (pomalidomide).

In some embodiments, the subject is further administered acorticosteroid.

In some embodiments, the corticosteroid is dexamethasone or predisone.

The second therapeutic agent or combinations thereof are typicallyadministered at dosages recommended for the agent.

The pharmaceutical composition of the invention may be administeredsimultaneously or sequentially with the second thereapeutic agent orcombinations thereof.

While having described the invention in general terms, the embodimentsof the invention will be further disclosed in the following examplesthat should not be construed as limiting the scope of the claims.

Example 1. Subcutaneous Delivery of 2% Human Immunoglobulin G (IgG) withRecombinant Human Hyaluronidase PH20 (rHuPH20) in the Miniature SwineModel Summary

The miniature pig is a preclinical model that is suitable for evaluatingsubcutaneous (SC) administration conditions of biotherapeutics due toits anatomical similarity to human skin and clinical translatability(Mahj et al., Exp Toxicol Path 57: 341-5, 2006). The objective of thisstudy was to assess and evaluate conditions for a 100 mL administrationof a 20 mg/mL human IgG solution containing 200, 500, or 800 U/mL ofrHuPH20 at two different flow rates (2 and 4 mL/min). Endpoints includedquantitative infusion pressure measurements as well as qualitativeassessments of the local infusion site, such as swelling size andfirmness.

Yucatan miniature pigs were subcutaneously infused with 100 mL of asolution containing 20 mg/mL immunoglobulin G (IgG) with 200, 500, or800 U/mL of rHuPH20 at a flow rate of 2 or 4 mL/min. Real-time in-linepressures were measured during infusions. Following the completion ofinfusions, local infusion sites were measured for visible swellingvolume and area if present, and qualitative scoring of the infusion sitefor presence and severity of erythema, size and firmness of theswelling/bleb, and gross observations. Infusion pressures were lowoverall and ranged from ˜40 to 60 mmHg (˜1 PSI) for both flow rates.

There were no statistical differences in pressure between the variousconcentrations of rHuPH20 and the two different flow rates, with overallpressure being slightly lower at the 2 mL/min flow rate as expected.

The unexpected finding was the number of infusions (10 of 12) that hadvisible and measurable swelling at the infusion site for the lower 2mL/min flow rate. This observation was observed at all threeconcentrations of rHuPH20. In contrast, only 3 of 12 infusions at thehigher 4 mL/min flow rate resulted in visible and measurable localswelling. Again this was observed at each concentration of rHuPH20.

In all cases where local swelling was visible, the swelling subsidedwithin an hour. Additionally, local swelling at the infusion sites wasgenerally soft to the touch and not indurated, as indicated by theswelling/induration index (mean score <2). The prevalence of erythemawas more frequent with infusions at the 2 mL/min flow rate; however,overall the severity of the erythema was mild and completely subsided bythe following day. No other gross observations of the infusion siteswere noted in the study.

Three different concentrations of rHuPH20 (200, 500, or 800 U/mL) wereevaluated in this study, with no statistical difference between theconcentrations based on the endpoints of the study. Overall, the higher4 mL/min flow rate resulted in lower frequency of erythema, visibleswelling, and local infusion site firmness than the 2 mL/min flow rate.

Test Articles and Methods

Test Articles

Materials in Formulation Buffer:

-   -   25 mM sodium acetate (Spectrum; PN #S0104; Lot #1DI0271)    -   60 mM sodium chloride (Spectrum; PN #50155; Lot #1CE0421)    -   140 mM D-mannitol (Spectrum; PN #MA165; Lot #1EB0316)    -   0.04% polysorbate 20 (JT Baker; PN #4116-04; Lot #0000017659)    -   pH 5.5 (glacial acetic acid to pH; Fisher Scientific; PN        #A491-212; Lot #080972)        Materials in Drug Substance:    -   Human Gamma Globulin (BioMed Supply; PN #HGG-1005; Lot        #BMS31309013)    -   rHuPH20 [(Manufactured by Cook Pharmica for Halozyme; Halozyme        Lot #462—    -   021B (revialed, Cook Lot #104-001-HSTFIL-9054)]        Formulation

20 mg/mL IgG was co-mixed with 200, 500, or 800 U/mL of rHuPH20 4 daysprior to study onset. The solutions were aliquoted into individual glassbottles, sealed with a stopper, and crimp capped. All solutions werestored at 2-8° C. until start of study, but were allowed to acclimate toroom temperature prior to infusions. Additionally, a sample was takenfrom each formulation for rHuPH20 enzymatic activity testing. Results ofthe enzymatic activity assay confirmed that all dosing solutions werewithin 10% of the target concentration (data not shown).

Animal Description

-   -   Species: Pig (Sus scrofa domestica)    -   Strain: Yucatan Miniature    -   Sex: Female    -   Age: >3 months    -   Weight: ˜12 kg    -   Quantity: 12    -   Source: S&S Farms (Ramona, Calif.)        Husbandry

Animals were housed in steel pens with automatic water provided adlibitum. Animals were fed twice daily (AM and PM), except on study day(PM only). Animal body weights were taken and recorded from day ofdelivery to one day post-completion of the study to assess animalhealth. All animals maintained body weights during this period (data notshown). The room environment was set to maintain a temperature of˜17-27° C. and a relative humidity of 40-70%, with a 12 hour light/12hour dark time cycle. Animals were allowed to acclimate to the facilityfor 7 days prior to study onset.

Test Materials

-   -   High pressure syringe pumps (KD Scientific; Holliston, Mass.)    -   23 ga×¾ inch winged infusion needle set with 12 inch tubing        (Terumo Medical Corporation; Somerset, N.J.)    -   140-cc Luer-lock syringe (Covidien; Mansfield, Mass.)    -   Extension set 7 inch (B/Braun; Bethlehem, Pa.)    -   PowerLab 4/30 (AD Instruments; Colorado Springs, Colo.)    -   Deltran-1 disposable pressure transducer (Utah Medical Products;        Midvale, Utah)    -   Digital caliper (Preisser Messtechnik; Gammertingen, Germany)    -   Isoflurane (Minrad International Company, Orchard Park, N.Y.)    -   Isoflurane vaporizer (VetEquip; Pleasanton, Calif.)        Experimental Design

The experimental design is summarized in the Description of Cohorts(Table 1) and Description of Infusions per Animal (Table 2). In brief,100 mL of a solution containing 20 mg/mL IgG co-mixed with 200, 500, or800 U/mL of rHuPH20 was administered into the abdominal region ofanesthetized Yucatan mini-pigs at a flow rate of 2 or 4 mL/min. Theendpoints of the study included infusion pressure measurements using anin-line pressure transducer, local post-infusion swelling (bleb) volumeand area (if possible), and qualitative assessment of the infusion site,including photographs.

TABLE 1 Cohort Test Article Flow Rate (mL/min) 1 IgG + 200 U/mL rHuPH202 mL/min 2 IgG + 500 U/mL rHuPH20 3 IgG + 800 U/mL rHuPH20 4 IgG + 200U/mL rHuPH20 4 mL/min 5 IgG + 500 U/mL rHuPH20 6 IgG + 800 U/mL rHuPH20

TABLE 2 Animal Flow rate ID (mL/min) Left side infusion Right sideinfusion 1 2 IgG + 200 U/mL IgG + 500 U/mL rHuPH20 rHuPH20 2 IgG + 200U/mL IgG + 800 U/mL rHuPH20 rHuPH20 3 IgG + 500 U/mL IgG + 200 U/mLrHuPH20 rHuPH20 4 IgG + 500 U/mL IgG + 800 U/mL rHuPH20 rHuPH20 5 IgG +800 U/mL IgG + 200 U/mL rHuPH20 rHuPH20 6 IgG + 800 U/mL IgG + 500 U/mLrHuPH20 rHuPH20 7 4 IgG + 200 U/mL IgG + 500 U/mL rHuPH20 rHuPH20 8IgG + 200 U/mL IgG + 800 U/mL rHuPH20 rHuPH20 9 IgG + 500 U/mL IgG + 200U/mL rHuPH20 rHuPH20 10 IgG + 500 U/mL IgG + 800 U/mL rHuPH20 rHuPH20 11IgG + 800 U/mL IgG + 200 U/mL rHuPH20 rHuPH20 12 IgG + 800 U/mL IgG +500 U/mL rHuPH20 rHuPH20 n = 4 infusions per cohort per flow rate Eachanimal received 2 different simultaneous infusions (one on eachcontralateral site) rHuPH20 concentrations: 200 U/mL = 20,000 total U;500 U/mL = 50,000 total U; 800 U/mL = 80,000 total U Infusion volume 100mL Endpoints: In-line (infusion) pressure Measurement of swelling/blebvolume and area Qualitative assessment of erythema and indurationincluding photographsStudy Procedure

Prior to start of study, animals were assessed for general health andbody weights were collected. On the day of study, animals wereanesthetized with isoflurane gas and placed in dorsal recumbence on aheated surgical table, and were maintained under isoflurane gas for theentire duration of the procedure. The abdominal region was cleaned withisopropanol and wiped dry with clean gauze. Infusion sites were locatedon the left and right abdominal regions, ˜3-4 cm towards the midlinestarting from the cranial end of the inguinal fold and then ˜6 cmcranial. The infusion sites were marked with a permanent marker and thenphotographed. Test articles were acclimated to room temperature prior toinfusions. Test articles were drawn into a 140 cc syringe (>100 mL toaccount for volume needed to prime the line). A pressure transducer wasattached to the syringe. A line extension set with an attached 23 ga×¾inch winged infusion needle was then attached to the transducer. Theinfusion hardware was then primed to the needle tip. The syringe wasloaded into the syringe pump. This process was done in duplicate, witheach syringe containing a different test article. The needles wereplaced subcutaneously into the marked left and right abdominal infusionsites of the animal. The in-line pressure transducer was zeroed. In-linepressure recordings were started and then the two syringe pumps werestarted simultaneously to infuse 100 mL of test articles at a flow rateof 2 or 4 mL/min. Upon completion of infusions, in-line pressure datacollection were stopped, needles removed, and the needle insertion holesealed with VetBond liquid adhesive to prevent any leakage. Localinfusion site swelling/bleb area and volume were measured using adigital caliper. Local infusion sites were also qualitatively assessedfor appearance (erythema), swelling/bleb size, and firmness (induration)using a 5 point scoring system (Table 3, Table 4, and Table 5,respectively). Finally, photographs were taken of the infusion sites.

TABLE 3 Scale Description 0 No erythema 1 Very slight erythema (barelyperceptible) 2 Well defined erythema 3 Moderate to severe erythema 4Severe erythema (beet redness) to slight eschar formation

TABLE 4 Scale Description 0 No swelling 1 Very slight swelling 2 Slightswelling 3 Moderate swelling 4 Severe swelling

TABLE 5 Scale Description 0 No perceptible difference in firmness afterinjection 1 Very slightly firm (barely perceptible) 2 Mildly firm 3Moderately firm 4 Very firmCalculations and Statistical MethodsAssessment of Infusion Pressure:

Infusion pressures, as measured via an in-line transducer, were recordedusing LabChart 7, and the mean pressure over the entire infusion periodwas calculated.

Assessment of Local Swelling Volume and Area:

Volume and area of post-infusion swelling were measured using a digitalcaliper and manually recorded. Measurements were recorded as length,width, and height. The formula for an ellipsoid was used to calculatevolume. Volume=4/3πABC, where A=radius of length, B=radius of width,C=radius of height. A simple formula of length×width was used tocalculate the area.

Assessment of Local Infusion Sites:

Local infusion sites were independently assessed by three separateevaluators following the completion of the infusion. Each evaluatorassessed the skin at each infusion site for the presence of erythema,size of local swelling, and firmness. A score on a 0 to 4 grading scalewas used to evaluate the three areas of assessment, with a score of 0representing no effect and 4 being severe. Additionally, erythema andswelling scores were used to calculate a primary irritation index (PII)using the formula PII=mean [(Σ of erythema grade+Σ of swellinggrade)÷2]. Furthermore, the swelling and firmness scores were used tocalculate a swelling/induration index (SII) using the formula SII=mean[(Σ of swelling grade+Σ of hardness grade)÷2]. A SII score of ≤2 was notconsidered to be indurated.

Statistical Analyses:

Statistical comparisons between cohorts were performed using an one-wayanalysis of variance (ANOVA) with a Tukey's multiple comparisons testfor continuous variables, and a nonparameteric Krusal-Wallis test with aDunn's multiple comparisons test for categorical variables. Statisticalsignificance was determined to be p<0.05.

Results

Assessment of Infusion Pressure:

100 mL of 20 mg/mL IgG co-mixed with 200, 500, or 800 U/mL of rHuPH20was administered into the abdominal region of Yucatan mini-pigs at aflow rate of 2 or 4 mL/min. Infusions at a flow rate of 2 mL/minresulted in mean infusion pressures of 40.5±0.1, 40.0±0.1, and 37.1±0.1mmHg±SEM for IgG co-mixed with 200, 500, and 800 U/mL of rHuPH20,respectively. Infusions at a flow rate of 4 mL/min resulted in meanpressures of 49.9±0.1, 55.5±0.1, and 61.9±0.2 mmHg±SEM for IgG co-mixedwith 200, 500, and 800 U/mL of rHuPH20, respectively. The infusionpressures were not statistically different between the variousconcentrations of rHuPH20 for each flow rate and were not statisticallydifferent between the two flow rates.

Assessment of Local Swelling Volume and Area:

After completion of each infusion, the local infusion site swelling wasmarked if visible and measured using a digital caliper. For infusions ata flow rate of 2 mL/min resulted in a mean swelling volume of 36.6±14.4,19.5±6.5, and 31.4±6.0 cm3±SEM for IgG co-mixed with 200, 500, and 800U/mL of rHuPH20, respectively, of which 10 of 12 infusions were visibleand measurable. On the contrary, the swelling volume was only detectedin 3 of 12 infusions at a flow rate of 4 mL/min, one at each rHuPH20concentration. The local swelling volumes were not statisticallydifferent between the various concentrations of rHuPH20 for each flowrate and were not statistically different between the two flow rates.

In addition to volume calculations, the local area of post-infusionvisible swelling was measured. For infusions at a flow rate of 2 mL/minresulted in a mean swelling area of 78.2±26.4, 59.7±20.0, and 94.9±9.7cm2±SEM for IgG co-mixed with 200, 500, and 800 U/mL of rHuPH20,respectively. For infusions at a flow rate of 4 mL/min resulted inmeasurable swelling areas in only 3 of 12 infusions, one at each rHuPH20concentration. The local swelling areas were not statistically differentbetween the various concentrations of rHuPH20 for each flow rate andwere not statistically different between the two flow rates.

Assessment of Local Infusion Sites:

Upon completion of infusions, the local infusion sites werequalitatively scored, by three separate evaluators, for presence andseverity of erythema, visible size of swelling, physical firmness of theskin, primary irritation index (PII) which incorporates the erythema andswelling scores, and swelling/induration index (SII) which incorporatesthe swelling and firmness scores to determine if induration was present.

The presence and severity of erythema was evaluated. For infusions at aflow rate of 2 mL/min resulted in a mean erythema score of (±SEM) of0.8±0.2, 0.4±0.1, and 1.0±0.3 for IgG commixed with 200, 500, and 800U/mL of rHuPH20, respectively. Infusions at a flow rate of 4 mL/minresulted in a mean erythema score (±SEM) of 0.3±0.1, 0.3±0.1, and0.2±0.1 for IgG co-mixed with 200, 500, and 800 U/mL of rHuPH20,respectively. The local erythema scores were not statistically differentbetween the various concentrations of rHuPH20 for each flow rate andwere not statistically different between the two flow rates. The size ofvisible local swelling was evaluated, and for infusions at a flow rateof 2 mL/min resulted in a mean swelling score (±SEM) of 1.9±0.4,1.4±0.3, and 2.0±0.2 for IgG co-mixed with 200, 500, and 800 U/mL ofrHuPH20, respectively. Infusions at a flow rate of 4 mL/min had fewervisible swelling with a mean erythema score (±SEM) of 0.6±0.3, 0.9±0.4,and 0.9±0.4 for IgG co-mixed with 200, 500, and 800 U/mL of rHuPH20,respectively. The local swelling scores were not statistically differentbetween the various concentrations of rHuPH20 for each flow rate and wasnot statistically different between the two flow rates, except forIgG+800 U/mL of rHuPH20 at 2 mL/min flow rate versus IgG+200 U/mL ofrHuPH20 at 4 mL/min flow rate (p<0.05). The physical firmness of theskin at the local infusion site was evaluated, and for infusions at aflow rate of 2 mL/min resulted in a mean firmness score (±SEM) of1.5±0.3, 1.0±0.2, and 1.4±0.2 for IgG co-mixed with 200, 500, and 800U/mL of rHuPH20, respectively. Infusions at a flow rate of 4 mL/min hadless local infusion site firmness with a mean firmness score (±SEM) of0.5±0.3, 0.7±0.2, and 0.7±0.3 for IgG co-mixed with 200, 500, and 800U/mL of rHuPH20, respectively. The local infusion site firmness scoreswere not statistically different between the various concentrations ofrHuPH20 for each flow rate and were not statistically different betweenthe two flow rates. The primary irritation index was calculated based onthe erythema and swelling scores. Infusions at 2 mL/min had a mean PIIscore (±SEM) of 1.4±0.3, 0.9±0.2, and 1.5±0.2 for IgG co-mixed with 200,500, and 800 U/mL of rHuPH20, respectively. Infusions at a flow rate of4 mL/min had lower PII scores with a mean score (±SEM) of 0.4±0.2,0.6±0.3, and 0.5±0.3 for IgG co-mixed with 200, 500, and 800 U/mL ofrHuPH20, respectively. The PII scores were not statistically differentbetween the various concentrations of rHuPH20 for each flow rate and wasnot statistically different between the two flow rates, except forIgG+800 U/mL of rHuPH20 at 2 mL/min flow rate versus IgG+200 U/mL ofrHuPH20 at 4 mL/min flow rate (p<0.05). The swelling/induration indexwas calculated based on the swelling and firmness scores. Infusions at 2mL/min had a mean SII score (±SEM) of 1.7±0.3, 1.2±0.2, and 1.7±0.2 forIgG co-mixed with 200, 500, and 800 U/mL of rHuPH20, respectively.Infusions at a flow rate of 4 mL/min had lower SII scores with a meanscore of 0.6±0.3, 0.8±0.3, and 0.8±0.3 (±SEM) for IgG co-mixed with 200,500, and 800 U/mL of rHuPH20, respectively. The SII scores were notstatistically different between the various concentrations of

rHuPH20 for each flow rate and was not statistically different betweenthe two flow rates. Based on the mean SII values (score of ≤2), thelocal infusions sites were considered not to be indurated. Lastly,photographs were taken prior to and after completion of each infusion.

Example 2. An Open-label, Multicenter, Dose Escalation Phase 1b Study toAssess the Safety and Pharmacokinetics of Subcutaneous Delivery ofDaratumumab with the Addition of Recombinant Human Hyaluronidase(rHuPH20) for the Treatment of Subjects with Relapsed or RefractoryMultiple Myeloma

The purpose of the study is to evaluate the safety, pharmacokinetics andantitumor activity of subcutaneous (SC) or intravenous (IV) delivery ofdaratumumab to participants with relapsed or refractory multiplemyeloma. This is an open-label multicenter, 2-part, Phase 1b doseescalation/expansion study to evaluate the safety, pharmacokinetics andantitumor activity of SC or IV delivery of daratumumab to participantwith relapsed or refractory multiple myeloma. Up to approximately 48participants in part 1 and 80 participants in part 2 will be enrolled.The Part 1 dose escalation phase is designed to determine therecommended Phase 2 dose (RP2D) based on safety and pharmacokinetic (PK)data of daratumumab. Each part of the study will have 3 phases: aScreening Phase, an open label treatment phase and a post-treatmentphase (from the final dose of study drug until post-treatment week 8).In Part 1, participants will be assigned to sequential cohorts ofapproximately 8 participants each cohort. Participants will be dosedwith DARA PH20 (Daratumumab with the Addition of Recombinant HumanHyaluronidase [rHuPH20]) by SC infusion once weekly in Cycles 1 (eachcycle 28 days) and 2, every 2 weeks in Cycles 3-6, and every 4 weeks insubsequent cycles of each cohort. After the last participant in eachcohort completes Cycle 3 day 1, the Safety Evaluation Team (SET) willevaluate the safety and pharmacokinetic data according toprotocol-defined criteria and make the decision whether to escalate thedose in a new cohort. The SET will review all safety and PK data fromPart 1 to determine the RP2D before initiation of Part 2. In Part 2,participants will be randomized 1:1 to receive recommended Phase 2 doseof DARA PH20 or IV delivery of 1200 mg DARA. Safety, pharmacokinetics,and antitumor activity of SC and IV delivery of daratumumab will beevaluated. Participant's safety will be monitored throughout the study.

Primary Outcome Measures:

Serum Trough Concentrations (Ctrough) of Daratumumab (Time Frame: Up topart 2 cycle 3 (each cycle 28 days) Day 1). Ctrough: the concentrationprior to study drug administration.

Part 1 and 2: Number of Participants with Adverse Events (AEs) andSerious AEs (Time Frame: Screening up to follow-up (30 days after lastdose administration)

An adverse event (AE) is any untoward medical occurrence in aparticipant who received study drug without regard to possibility ofcausal relationship. A serious adverse event (SAE) is an AE resulting inany of the following outcomes or deemed significant for any otherreason: death; initial or prolonged inpatient hospitalization;life-threatening experience (immediate risk of dying); persistent orsignificant disability/incapacity; congenital anomaly.

Secondary Outcome Measures:

-   -   Part 1 and 2: Serum Concentration of Daratumumab and Recombinant        Human Hyaluronidase (rHuPH20) (Plasma) Antibodies (Time Frame:        Approximately 2 years). Serum levels of antibodies to        Daratumumab and rHuPH20 for evaluation of potential        immunogenicity.    -   Part 1 and 2: Percentage of Participants with Complete Response        (CR) (Time Frame: Approximately 2 years). CR is Defined as the        proportion of Participants achieving CR (including sCR)        according to the International Myeloma Working Group (IMWG)        criteria.    -   Part 1 and 2: Percentage of Participants With Overall Response        Rate (ORR) (Time Frame: Approximately 2 years). Overall response        rate is defined as the percentage of participants who achieve        complete response or partial response according to the        International Myeloma Working Group criteria, during or after        study treatment.    -   Part 1 and 2: Duration of Response (DR) (Time Frame:        Approximately 2 years). The DR is time from date of initial        documentation of response (CR or PR) to date of first documented        PD, as defined by IMWG criteria.    -   Part 1 and 2: Time to Response (Time Frame: Approximately 2        years). Time to response is defined as the time from the date of        first dose of study treatment to the date of the first        documentation of observed response (CR or PR).

Table 6 shows the study design. Table 7 shows the interventions.

TABLE 6 Arm Number or Name Type Description Part 1: Cohort 1Experimental Participants will receive DARAPH20, 1200 mg (daratumumab1200 milligram (mg) with Recombinant Human Hyaluronidase [rHuPH20]30,000 U) by Subcutaneous (SC) infusion once weekly in Cycles 1 (eachcycle is 28 days) and 2, every 2 weeks in Cycles 3-6, and then every 4weeks in subsequent cycles until disease progression. Part 1: Cohort 2Experimental Participants will receive DARAPH20, 1800 mg (daratumumab1800 milligram (mg) with Recombinant Human Hyaluronidase [rHuPH20]45,000 U) by SC infusion once weekly in Cycles 1 and 2, every 2 weeks inCycles 3-6, and then every 4 weeks in subsequent cycles until diseaseprogression. Part 1: Cohort 3 Experimental Participants will receiveDARAPH20 at a dose which will be decide by Study Evaluation Team (SET)once weekly by SC infusion in Cycles 1 and 2, every 2 weeks in Cycles3-6, and then every 4 weeks in subsequent cycles until diseaseprogression. Also up to three additional optional cohorts (Cohorts 3b,3c, and 3d) may be enrolled to repeat a dose level of daratumumab Part2: Cohort 4 Experimental DARA-PH20 SC infusion at the RP2D (recommendedPhase 2 dose) that is identified in Part 1 will be administered toparticipants by Sc infusion once weekly in Cycles 1 and 2, every 2 weeksin Cycles 3-6, and then every 4 weeks in subsequent cycles until diseaseprogression. Part 2: Cohort 5 Experimental Daratumumab 1200 mg will beadministered to participants by intravenous (IV) once weekly in Cycles 1and 2, every 2 weeks in Cycles 3-6, and then every 4 weeks in subsequentcycles until disease progression.

TABLE 7 Intervention Name Type Associated Arms Description DaratumumabDrug Part 1: Cohort 1 Participants will Subcutaneous Part 1: Cohort 2receive Daratumumab (SC) Part 1: Cohort 3 sc infusion once weeklyinfusion Part 2: Cohort 4 in Cycles 1 (each cycle is 28 days) and 2,every 2 weeks in Cycles 3-6, and every 4 weeks in subsequent cycles.Recombinant Part 1: Cohort 1 Participants will Human Part 1: Cohort 2receive Recombinant Hyaluronidase Part 1: Cohort 3 Human Hyaluronidase[rHuPH20]) SC Part 2: Cohort 4 [rHuPH20]) along with infusionDaratumumab by SC infusion once weekly in Cycles 1 (each cycle is 28days) and 2, every 2 weeks in Cycles 3-6, and every 4 weeks inDaratumumab Part 2: Cohort 5 Participants will Intravenous (IV) receiveDaratumumab infusion 1200 mg IV infusion once weekly in Cycles 1 (eachcycle 28 days) and 2, every 2 weeks in Cycles 3-6, and every 4 weeks insubsequent cycles.Eligibility

Participants proven to have symptomatic (having symptoms) multiplemyeloma (MM) according the International Myeloma Working Group (IMWG)diagnostic

Criteria:

-   -   Measurable disease as defined by any of the following: (a)        immunoglobulin (Ig) G myeloma (serum monoclonalparaprotein        [M-protein] level>=1.0 gram/deciliter [g/dL] or urine M-protein        level greater than or equal to (>=) 200 milligram[mg]/24        hours[hrs]; or (b) IgA, IgD, or IgE multiple myeloma (serum        M-protein level>=0.5 g/dL or urine Mprotein level>=200 mg/24        hrs); or (c) light chain multiple myeloma (serum immunoglobulin        free light chain>=10 mg/dL and abnormal serum immunoglobulin        kappa lambda free light chain ratio)    -   Participant must have an Eastern Cooperative Oncology Group        (ECOG) performance status score of 0, 1, or 2    -   Pretreatment clinical laboratory values must meet        protocoldefined parameters during the Screening phase    -   Man, who is sexually active with a woman of child-bearing        potential and has not had a vasectomy, must agree to use an        adequate contraception method as deemed appropriate by the        Investigator, and must also agree to not donate sperm during the        study and 4 months after last dose of daratumumab        Exclusion Criteria:    -   Participant has received daratumumab or other anti-cluster of        differentiation 38 (anti-CD38) therapies previously    -   Participant has received anti-myeloma treatment within 2 weeks        before Cycle 1 Day 1    -   Participant has previously received an allogenic stem cell        transplant; or participant has received autologous stem cell        transplantation (ASCT) within 12 weeks before Cycle 1 Day 1    -   Participant has a history of malignancy (other than multiple        myeloma) within 5 years before Cycle 1 Day 1 (exceptions are        squamous and basal cell carcinomas of the skin and carcinoma in        situ of the cervix, or malignancy that in the opinion of the        Investigator, with concurrence with the Sponsor's medical        monitor, is considered cured with minimal risk of recurrence)    -   Participant is exhibiting clinical signs of meningeal        involvement of multiple myeloma    -   Gender: Both    -   Age limit: 18 years    -   Accepts healthy volunteers: No        Interim Readout of Part 1 (Data Cutoff 21 Jul. 2016 for        Safety/Demographics/Disease History and 28 Jul. 2016 for the        Efficacy Data)

Methods

Patients had RRMM with ≥2 prior lines of therapy including a proteasomeinhibitor (PI) and an immunomodulatory drug (IMiD). Part 1 of the 2 partstudy enrolled sequential cohorts at 1200 mg and 1800 mg DARA doselevels to determine the recommended SC dose for Part 2. DARA-PH20 wasadministered in 4-week treatment cycles: QW for 8 weeks, Q2W for 16weeks, and Q4W thereafter. DARA-PH20 was infused in 1200 mg doses in 60mL over 20 min or 1800 mg in 90 mL over 30 min, via a syringe pump atrotating sites on the abdomen. Pre- and/or post-infusion medicationsincluded paracetamol, diphenhydramine, montelukast, andmethylprednisolone. In part 2, pts will be randomized 1:1 to receive therecommended phase 2 dose (RP2D) of SC DARA-PH20 or IV DARA (16 mg/kg).The RP2D of DARA-PH20 will be selected based on a cumulative review ofthe pharmacokinetic and safety data obtained from part 1 and shouldachieve a maximum serum Ctrough during weekly dosing that is similar toor higher than that observed for the approved 16 mg/kg IV dose. Primaryendpoints were Ctrough of DARA up to Cycle 3 Day 1 and safety. Secondaryendpoints included overall response rate (ORR).

Results

To date, 41 pts were treated in part 1 with SC DARA-PH20 at the 1200 mg(n=8) and 1800 mg (n=33) dose levels. Infusion related reactions (IRRs)were reported in 9/41 pts (22%) and were mostly grade ½ in severityincluding chills, fever, rigors, vomiting, itching, edema of the tongue,non-cardiac chest pain and wheezing. One pt developed grade 3 dyspneaand 1 pt required hospitalization due to fever and chills (both grade 2)after the first infusion. All IRRs developed during or within 6 hours ofthe first SC infusion and were controlled with antihistamine,corticosteroid, antiemetic, or bronchodilator treatment. No IRRs werereported with subsequent infusions. Overall, the adverse event profileof DARA-PH20 was consistent with that of IV DARA. Grade 3 or higherdrug-related adverse events were reported in 5/41 (12%) pts includingfatigue (2 pts), influenza, hypertension, dyspnea, and tumor lysissyndrome. SC administration of DARA-PH20 was well tolerated at theabdominal wall injection site with 3/41 (7%) pts reporting grade 1erythema, induration, or burning sensation. Analysis showed a higher maxCtrough in the 1800 mg cohort in comparison to the max Ctrough achievedfollowing IV DARA (16 mg/kg).

In the 1200 mg cohort of 8 pts (median of 5 lines of prior therapy[range 2-10]; prior ASCT, 63%; PI refractory only, 0%; IMiD refractoryonly, 13%; double refractory to PI and IMiD, 63%) a 25% ORR was observedincluding 2 partial responses (PR). Median time to response was 14(range 8-20) weeks. Among 17 response evaluable pts in the 1800 mgcohort with cycle 3 day 1 assessments (median of 4 prior lines oftherapy [[range 2-7]; prior ASCT, 76%; PI refractory only, 6%; IMiDrefractory only, 12%; double refractory to PI and IMiD, 65%) a 41% ORRwas observed consisting of 3 very good partial responses and 4 PRs.Median time to response was 4 (range 4-8) weeks.

Conclusions:

SC DARA-PH20 was well tolerated and achieved serum trough concentrationssimilar to or greater than IV DARA with a lower rate of IRRs compared toIV DARA over a significantly shorter infusion time. Preliminary datasuggest that in this pt population SC DARA-PH20 may enable similarresponse rates to IV DARA monotherapy. The 1800 mg dose level ofDARA-PH20 was selected as the RP2D for part 2 of the study. These earlydata support further study of SC DARA in clinical trials.

Example 3. Development of Co-Formulations of Daratumumab andHyaluronidase

Various co-formulations were evaluated in order to establish the overallphysico-chemical stability and delivery of daratumumab and rHuPH20 inthe co-formulated product. The impact of the concentrations of theactive constituent and/or the excipients in the formulations wasevaluated in some of the stability and/or animal studies (shelfstability, shaking stability and in pig infusion studies). Table 8provides a summary of the formulations that have been used in variousstudies.

TABLE 8 Sorbitol/ Daratumumab rHuPH20 His Sucrose PS20 Met Formulation(mg/mL) (U/mL) (mM) (mM) (% w/v) (mg/mL) pH 1 100 500 10 300 0.04 2 5.52 120 2000 10 300 0.04 1 5.6 3 100 500 10 300 0.0  2 5.5 4 100 500 10300 0.01 2 5.5 5 100 500 10 300 0.02 2 5.5 6 100 500 10 300 0.06 2 5.5 7100 0 10 200/ 100 0.04 0 5.5 8 100 0 10 100/ 200 0.04 0 5.5 9 100 50 10300 0.04 1 5.5 10 100 500 10 300 0.04 1 5.5 11 100 2000 10 300 0.04 15.5 12 100 5000 10 300 0.04 1 5.5 His: histidine Met: methionine

The ranges of the excipients and the active constituents in the testedformulations are shown in Table 9.

TABLE 9 Formulation component Range rHuPH20 0-2000 U/mL Daratumumab100-120 mg/mL Histidine 10 mM Sorbitol 100-300 mM Sucrose 0-200 mMPolysorbate-20 (PS20) 0.0-0.04% (w/v) Methionine 0-2 mg/mL

The generated formulations were tested in various assays for theircharacteristics, including evaluation of sub-visible particles, microflow imaging (WI), size exclusion chromatography (SEC), capillaryiso-electric focusing (cIEF), SDS-PAGE (non-reducing and reducing),peptide mapping, extractable volume, turbidity, osmolality, and pH.

Sub-visible particles (Sub-vis): Number of sub-visible particles sizesof ≥10 μm or ≥25 μm is usually aggregates of protein molecules and canbe assayed by the light obscuration HIAC method whereby the solution ispassed through a small orifice and the blockage of light provides theinformation on the particle size passing through.

MFI: An orthogonal to the light obscuration method, micro flow imaging(MFI) takes snapshot images of particles flowing through and re-convertsback to the number of particles present in a particular volume ofliquid. This method provides information about the large aggregates ofproteins present in the solution.

SEC: A size exclusion chromatographic separation method whereby a columnis used to distribute the molecules within the solution flowing throughaccording to their broad size range. Monomers, aggregates and fragmentselute at different times from the column and hence their relativeproportions in a sample can be quantified using a standard UV detector.

cIEF: Capillary iso-electric focusing distributes the moleculesaccording to the charge on the molecule and is a good indicator of theoverall chemical stability. For example deamidation may result in achange in the charge of the molecule and thus would be picked up by thismethod. The method provides an idea of the total acidic, basic andintact % of molecules present in the solution.

SDS (reducing and non-reducing conditions): SDS method providesinformation on the physical stability of the molecule. SDS provides ameasure of the intact, aggregated and fragmented species present in thesolution. Non reducing SDS provides information on the respectiveintact, aggregated and fragmented constituents of the antibody whilereducing SDS (after disulfide disruption) provides the same informationfor the heavy and light chains of the antibody.

Peptide mapping: Peptide mapping is an essential technique for studyingthe primary structure of proteins. For recombinant proteinpharmaceuticals, peptide mapping is used for the initial proof ofstructure characterization. Peptide mapping also provides information onpost translational modifications such as deamidation, oxidation etc

Extractable volume: The method provides information on the amount/volumeof liquid that can be withdrawn from the vial after the respective timepoint.

Turbidity: A light scattering based method to evaluate the physicalstability of the solution. An increase in the size of the particles oraggregates results in an increase in the light scattering signal and ishence picked up as turbidity (opalescence) of the solution. Turbidity ismeasured in Nephelometric Turbidity Units (NTU).

Osmolality: Provides a measure of the total osmotic activity which isdependent on the total true activity of the molecules (activitycoefficient multiplied by concentration). The solution must be close tothe osmolality of the serum to be injectable.

pH: Provides an idea of the overall stability and is important that thepH remains constant throughout the shelf life.

rHuPH20 enzymatic activity: The determination of hyaluronidase activityis based on the formation of a precipitate when hyaluronic acid (HA)binds with acidified serum. The activity is measured by incubatinghyaluronidase with HA for 30 minutes in a 96-well plate format at 37° C.and then precipitating the undigested HA with the addition of acidifiedserum. The resulting turbidity is measured at 640 nm and the decrease inturbidity resulting from enzymatic cleavage of the HA substrate is ameasure of the hyaluronidase activity.

Shelf stability of Formulation 1 (100 mg/mL Daratumumab, 10 mMHistidine, 300 mM Sorbitol, 0.04% PS20, 2 mg/mL Methionine, 500 U/mLPrHuh20, pH 5.5) was evaluated using assays described. Samples were puton stability in 25R vials (filled at 16 mL volume) at differenttemperatures (5, 25 and 40° C.) and vials were pulled for analysis usingvarious assays at different time points (0, 1, 2, 3, 4, 5 and/or 6months). Data indicates that the co-formulated product is stable underthe storage conditions both with respect to the Daratumumab as well asrHuPH20 as indicated by various assays. The profile as observed forparticles, color, turbidity, sec etc was very similar to well behavedstable antibodies and the data is comparable to the stability data ofsome commercial mAb formulations.

Table 10 shows the number of particles in Formulation 1 over time asassessed using HIAC.

Table 11 shows the number of particles in Formulation 1 over time asassessed using MFI.

Table 12 shows the pH of Formulation 1 over time.

Table 13 shows the turbidity of Formulation 1 over time.

Table 14 shows the proportion of high-molecular weight aggregates andlow molecular weight fragments in Formulation 1 over time.

Table 15 shows the acidic and basic species in Formulation 1 over timeas assessed using cIEF.

Table 16 shows the percent (%) purity of Formulation 1 over time asassessed using reduced SDS-PAGE.

Table 17 shows the percent (%) purity of Formulation 1 over time asassessed using non-reduced SDS-PAGE.

Table 18 shows the percent (%) bioactivity of daratumumab and enzymeactivity of rhPH20 in Formulation 1 over time.

TABLE 10 Average Cumulative Count / mL of Storage particle sizes ≥10 μMor ≥25 μM Temperature 0 months 3 months 6 months (° C.) ≥10 μm ≥25 μm≥10 μm ≥25 μm ≥10 μm ≥25 μm  5 7.17 6.33 6.5 4.8 42.50 30.83 25 7.176.33 5.5 1.8 110.50 81.83 40 7.17 6.33 64.5 44.8

TABLE 11 0 3 6 months months months Temperature 5 5 25 40 5 25 (° C.)Particles/mL 539 1155 3668 3371 686 3581 ≥2-<10 μm ECD Particles/mL 9.339 93 80 36 105 ≥10-<25 μm ECD Particles/mL 1.4 2.6 13 9.3 7.4 7.2≥25-<70 μm ECD Particles/mL 0.5 0.2 0.2 0.2 0.0 0.5 ≥70 μm ECD

TABLE 12 Storage Temperature pH over Storage Time (months) (° C.) 0 m 1m 2 m 3 m 6 m 5 5.7 NA 5.7 5.7 5.6 25 5.7 5.7 5.7 5.7 5.6 40 5.7 5.7 5.65.7 NA m: month NA: not analyzed

TABLE 13 Storage Temperature Average NTU over Storage Time (months) (°C.) 0 m 1 m 2 m 3 m 6 m 5 3.5 3.5 3.5 3.4 25 3.5 3.5 3.6 3.7 4.6 40 3.54 5.5 8.2 NA m: month NTU: Nephelometric Turbidity Units NA: notanalyzed

TABLE 14 Storage Storage Time Temperature Percentage (%) of species(months) (° C.) HMW Monomer LMW 0 5 0.74 99.25 0.01 25 0.74 99.25 0.0140 0.74 99.25 0.01 3 5 0.84 99.16 0.02 25 1.11 98.75 0.14 40 1.87 94.503.64 6 5 0.89 99.10 NA 25 1.30 98.38 NA 40 NA NA NA HMW: high molecularweight species LMW: low molecular weight species NA: not analyzed

TABLE 15 Storage Storage % % % Time Temperature Main Acidic Basic(months) (° C.) peak peaks peaks 0 5 69.2 27.8 3 25 69.2 27.8 3 40 69.227.8 3 3 5 68.2 28.9 2.9 25 63 32.8 4.2 40 31.3 60.2 8.5 6 5 68.1 28.73.2 25 55.4 38 6.6 40 NA NA NA NA: not analyzed

TABLE 16 Storage Temperature % Purity over Storage Time (in months) (°C.) 0 m 1 m 2 m 3 m 6 m 5 98.16 NA 97.96 98 98.02 25 98.16 97.87 97.6697.57 96.75 40 98.16 96.4 95 92.88 NA m: month NA: not analyzed

TABLE 17 Storage Temperature % Purity over Storage Time (in months) (°C.) 0 1 2 3 6 5 97.56 97.64 97.64 97.5 25 97.56 97.27 96.97 96.62 95.6340 97.56 94.84 92.61 89.37 NA m: month NA: not analyzed

TABLE 18 Storage Time and Temperature 0 6 months 6 months Molecule Assaymonths 5° C. 25° C. Daratumumab ADCC* 102 103 83 CDC* 95 101 93 PH20Enzymatic 574 600 or 609 activity** 584? *Percent control **U/mL

Agitation (shaking) stability of the Formulation 1 was also assessedusing the above assays to characterize the formulations and study theimpact of PS concentrations by varying just PS20 concentrations in thatformulation (Formulations 1, 3, 4, 5 and 6 in Table 8 where PS20 wasvaried to 0, 0.01, 0.02, 0.04, 0.06%). The data indicated that theco-formulation was stable under the shaking conditions both with respectto the Daratumumab as well as the enzyme as indicated by various assays.The profile as observed for particles, color, turbidity, sec etc wasvery similar to well behaved stable antibodies for all concentrations ofPS but 0% (0% PS20 formulation had particles and was not stable) and thedata was comparable to the stability data of some commercial mAbformulations (data not shown).

Shelf stability of Formulation 2 (120 mg/mL daratumumab, 10 mMHistidine, 300 mM Sorbitol, 0.04% PS20, 1 mg/mL Methionine, 2000 U/mLrhuPH20, pH 5.5) was evaluated using assays described. Samples were puton stability in 25R vials filled at 13.27 mL volume with overfill (1500mg dose) at different temperatures and vials were pulled for analysisusing various assays as below. The collected data indicated that theco-formulated product is stable under the storage conditions both withrespect to the daratumumab as well rHuPH20. The profile as observed forparticles, color, turbidity, sec etc was very similar to well behavedstable antibodies and the data was comparable to the stability data ofsome commercial mAb formulations. rhuPH20 is very susceptible at highertemperatures and loses all activity very fast when stored at 40° C.Table 19 shows the characteristics of the formulation.

TABLE 19 Storage Time, Temperature and relative humidity (RH) 0 1 month1 month months 40° C./ 25° C./ Characteristics and/or assay 5° C. 75% RH60% RH Average Cumulative Count/mL 58.89 375.68 4285.81 of particlesizes 2-10 μm Average Cumulative Count/mL 3.34 12.96 1.98 of particlesizes 10-25 μm Average Cumulative Count/mL 1.23 .49 .12 of particlesizes ≥25 μm PS20 (% w/v) 0.038 0.02 0.03 pH 5.6 5.6 5.6 Turbidity (NTU)5 11 6 % Purity (cSDS, reducing) 98.4 98.6 98.1 % AGHC, aglycosylatedheavy 0.4 0.5 0.4 chain (cSDS, reducing) % Purity (cSDS, non-reducing)97.7 95.2 97.7 % monomer, SE-HPLC 99.1 98.0 98.8 % aggregate, SE-HPLC0.9 1.6 1.1 % fragments SE-HPLC <0.10 0.4 <0.10 Daratumumab bioactivity,CDC 105 88 99 (% control) Daratumumab bioactivity, ADCC 99 73 103 (%control) rhuPH20 activity (U/mL) 2205 0 2258

Formulations 3-8 were tested for their shelf stability or shakingstability using some or all assays described. The data indicated thatthe Formulations 3-8 were stable under the conditions assessed both withrespect to the daratumumab as well as HuPH20 (formulations 7 and 8 hadno rHuPH20). Methionine was included into formulations 1-6 and 9-12 toprovide added oxidation stability. The profile as observed forparticles, color, turbidity, sec etc was very similar to well behavedstable antibodies and the data was comparable to the stability data ofsome commercial mAb formulations (data not shown).

Agitation (shaking) stability of the Formulation 1 was also assessedusing the above assays to characterize the formulations and study theimpact of PS20 concentrations by varying just PS20 concentrations inthat formulation (Formulations 1, 3, 4, 5 and 6 in table 8 where PS20was varied to 0, 0.01, 0.02, 0.04, 0.06%). The data indicated that theco-formulation was stable under the shaking conditions both with respectto daratumumab as well as rHuPH20 as indicated by various assays. Theprofile as observed for particles, color, turbidity, sec etc was verysimilar to well behaved stable antibodies for all concentrations of PSbut 0% and the data was comparable to the stability data of somecommercial mAb formulations (data not shown).

Formulations 9-12 were also evaluated for the assessment of subcutaneousadministration of daratumumab with varying concentrations of enzyme in aswine model as described in Example 2. These studies were conducted todetermine a suitable concentration of rhPH20 to deliver 16 mL ofdaratumumab. End points were infusion pressure, area of swelling or blebif measurable and qualitative assessment of site. Dose dependentincrease in infusion pressure was seen. All rhPH20 concentrations tested(50, 500, 2000, 5000 U/mL were sufficient to deliver 16 mL ofdaratumumab.

We claim:
 1. A pharmaceutical composition comprising an anti-CD38antibody at a concentration of about 120 mg/mL and a hyaluronidase at aconcentration of about 2000 U/mL, wherein the anti-CD38 antibody is ofan IgG1 isotype and comprises a heavy chain variable region sequence ofSEQ ID NO: 4 and a light chain variable region sequence of SEQ ID NO: 5,and wherein the hyaluronidase is rHuPH20 recombinant hyaluronidase. 2.The pharmaceutical composition of claim 1, further comprising apharmaceutically acceptable carrier.
 3. The pharmaceutical compositionof claim 1, wherein the anti-CD38 antibody comprises a heavy chainsequence of SEQ ID NO: 12 and a light chain sequence of SEQ ID NO: 13.4. The pharmaceutical composition of claim 2, further comprising a) fromabout 5 mM to about 50 mM histidine; and b) from about 50 mM to about400 mM sorbitol.
 5. The pharmaceutical composition of claim 4, furthercomprising a) from about 0.01% w/v to about 0.1% w/v polysorbate-20(PS-20); b) from about 0.1 mg/mL to about 2.5 mg/mL methionine; or c)from about 0.01% w/v to about 0.1% w/v PS-20 and from about 0.1 mg/mL toabout 2.5 mg/mL methionine.
 6. The pharmaceutical composition of claim4, comprising a) about 10 mM histidine; and b) from about 100 mM toabout 300 mM sorbitol.
 7. The pharmaceutical composition of claim 6,further comprising a) from about 0.01% w/v to about 0.04% w/vpolysorbate-20 (PS-20); and b) from about 1 mg/mL to about 2 mg/mLmethionine.
 8. The pharmaceutical composition of claim 5, comprising a)about 10 mM histidine; b) about 300 mM sorbitol; c) about 0.04% w/vPS-20; and d) about 1 mg/mL methionine; pH about 5.6.
 9. Thepharmaceutical composition of claim 5, comprising a) about 10 mMhistidine; b) about 300 mM sorbitol; c) about 0.04% w/v PS-20; and d)about 2 mg/mL methionine; pH about 5.5.
 10. The pharmaceuticalcomposition of claim 5, comprising a) about 10 mM histidine; b) about300 mM sorbitol; c) about 0.01% w/v PS-20; and d) about 2 mg/mLmethionine; pH about 5.5.
 11. The pharmaceutical composition of claim 5,comprising a) about 10 mM histidine; b) about 300 mM sorbitol; c) about0.02% w/v PS-20; and d) about 2 mg/mL methionine; pH about 5.5.
 12. Thepharmaceutical composition of claim 5, comprising a) about 10 mMhistidine; b) about 300 mM sorbitol; c) about 0.06% w/v PS-20; and d)about 2 mg/mL methionine; pH about 5.5.
 13. The pharmaceuticalcomposition of claim 5, comprising a) about 10 mM histidine; b) about300 mM sorbitol; c) about 0.04% w/v PS-20; and d) about 1 mg/mLmethionine; pH about 5.5.
 14. The pharmaceutical composition of claim 2which is in a unit dosage form.
 15. A method of treating a CD38-positivemultiple myeloma (MM) in a subject, comprising administeringsubcutaneously to the subject in need thereof a pharmaceuticalcomposition comprising an anti-CD38 antibody at a concentration of about120 mg/ml and a hyaluronidase at a concentration of about 2000 U/ml, fora time sufficient to treat the CD38-positive MM, wherein the anti-CD38antibody is of an IgG1 isotype and comprises a heavy chain variableregion sequence of SEQ ID NO: 4 and a light chain variable regionsequence of SEQ ID NO: 5, and wherein the hyaluronidase is rHuPH20recombinant hyaluronidase.
 16. The method of claim 15, wherein theanti-CD38 antibody comprises a heavy chain sequence of SEQ ID NO: 12 anda light chain sequence of SEQ ID NO:
 13. 17. The method of claim 15,wherein the pharmaceutical composition further comprises from about 5 mMto about 50 mM histidine and from about 50 mM to about 400 mM sorbitol,and optionally further comprises from about 0.01% w/v to about 0.1% w/vpolysorbate-20 (PS-20), from about 0.1 mg/mL to about 2.5 mg/mLmethionine, or from about 0.01% w/v to about 0.1% w/v PS-20 and fromabout 0.1 mg/mL to about 2.5 mg/mL methionine.
 18. The method of claim17, wherein the pharmaceutical composition comprises about 10 mMhistidine and from about 100 mM to about 300 mM sorbitol, optionallyfurther comprising from about 0.01% w/v to about 0.04% w/v PS-20, fromabout 1 mg/mL to about 2 mg/mL methionine, or from about 0.01% w/v toabout 0.04% w/v PS-20 and from about 1 mg/mL to about 2 mg/mLmethionine.
 19. The method of claim 18, wherein the pharmaceuticalcomposition comprises about 10 mM histidine, about 300 mM sorbitol,about 0.04% w/v PS-20 and about 1 mg/mL methionine; pH about 5.6. 20.The method of claim 18, wherein the pharmaceutical composition comprisesabout 10 mM histidine, about 300 mM sorbitol, about 0.04% w/v PS-20 andabout 2 mg/mL methionine; pH about 5.5.
 21. The method of claim 15,further comprising administering a second therapeutic agent.
 22. Themethod of claim 21, wherein the second therapeutic agent is a proteasomeinhibitor, an alkylating agent or a glutamic acid derivative, orcombinations thereof.
 23. The method of claim 22, wherein a) theproteasome inhibitor is bortezomib, carfilzomib or ixazomib; b) thealkylating agent is busulfan, cyclophosphamide, bendamustine,chlorambucil, carboplatin, cisplatin, temozolomide, melphalan,carmustine, lomustine, dacarbazine, oxaliplatin, ifosfamide,mechlorethamine, thiotepa, trabectedin or streptozocin; and c) theglutamic acid derivative is lenalidomide, thalidomide or pomalidomide.24. The method of claim 23, further comprising administering acorticosteroid.
 25. The method of claim 24, wherein the corticosteroidis dexamethasone or prednisone.
 26. The method of claim 25, wherein thecorticosteroid is dexamethasone.
 27. A unit dosage form, comprising a)an anti-CD38 antibody at a concentration of about 120 mg/mL, wherein theanti-CD38 antibody is of an IgG1 isotype and comprises a heavy chainvariable region sequence of SEQ ID NO: 4 and a light chain variableregion sequence of SEQ ID NO: 5; b) a hyaluronidase at a concentrationof about 2000 U/mL, wherein the hyaluronidase is rHuPH20 recombinanthyaluronidase; c) histidine at a concentration of from about 5 mM toabout 15 mM; d) sorbitol at a concentration of from about 100 mM toabout 300 mM; e) polysorbate-20 (PS-20) at a concentration of from about0.01% w/v to about 0.04% w/v; and f) methionine at a concentration offrom about 1 mg/mL to about 2 mg/mL, at a pH of about 5.5.
 28. The unitdosage form of claim 27, wherein histidine is present at a concentrationof about 10 mM.
 29. The unit dosage form of claim 28, wherein sorbitolis present at a concentration of about 300 mM.
 30. The unit dosage formof claim 29, wherein polysorbate is present at a concentration of about0.04% w/v.
 31. The unit dosage form of claim 30, wherein methionine ispresent at a concentration of about 1 mg/mL.
 32. The unit dosage form ofclaim 31, comprising sucrose at a concentration of from about 100 mM toabout 200 mM.
 33. The unit dosage form of claim 32, wherein theanti-CD38 antibody comprises a heavy chain sequence of SEQ ID NO: 12 anda light chain sequence of SEQ ID NO:
 13. 34. A container comprising theunit dosage form of claim
 33. 35. A unit dosage form, comprising a) ananti-CD38 antibody at a concentration of about 120 mg/mL, wherein theanti-CD38 antibody is of an IgG1 isotype and comprises a heavy chainvariable region sequence of SEQ ID NO: 4 and a light chain variableregion sequence of SEQ ID NO: 5; b) a hyaluronidase at a concentrationof about 2000 U/mL, wherein the hyaluronidase is rHuPH20 recombinanthyaluronidase; c) histidine at a concentration of about 10 mM; d)sorbitol at a concentration of about 300 mM; e) polysorbate-20 (PS-20)at a concentration of about 0.04% w/v; and f) methionine at aconcentration of about 1 mg/ml, at a pH of about 5.6.
 36. A containercomprising the unit dosage form of claim
 35. 37. The unit dosage form ofclaim 35, wherein the anti-CD38 antibody comprises a heavy chainsequence of SEQ ID NO: 12 and a light chain sequence of SEQ ID NO: 13.